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PCR Enzymes TransFastTaqDNA Polymerase
EasyTaq
DNA Polymerase
EasyTaq
DNA Polymerase for PAGE
TransTaq-T DNA Polymerase
TransTaq
DNA Polymerase High Fidelity (HiFi)
TransStartTaqDNA Polymerase
TransStartTopTaqDNA Polymerase
EasyPfuDNA Polymerase TransStart
FastPfuDNA Polymerase
TransStartFastPfuFly DNA Polymerase
GC Enhancer
PCR Stimulant
PCR SuperMix
2EasyTaq
PCR SuperMix
2EasyTaqPCR SuperMix for PAGE
2TransTaq-T PCR SuperMix
2TransTaq High Fidelity (HiFi) PCR SuperMix
2EasyPfuPCR SuperMix
2TransStartFastPfuPCR SuperMix
Direct PCR
TransDirectTM
Animal Tissue PCR Kit
TransDirectTM Plant Tissue PCR Kit
TransDirectTM
Blood PCR Kit
RT-PCR
EasyScript Reverse Transcriptase
TransScript
Reverse Transcriptase
TransScript II Reverse Transcriptase
EasyScript
First-Strand cDNA Synthesis SuperMix
TransScript First-Strand cDNA Synthesis SuperMix
TransScript
One-Step gDNA Removal and cDNA
Synthesis SuperMix
005
006
007
009
010
012
014
016017
019
021
022
024
025
026
Chapter 1 PCR, RT-PCR,qPCR and qRT-PCR
027
029
030
031
033
034
038
040
041
042
044
045
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qPCR and qRT-PCR SuperMix
TransStart
Green qPCR SuperMix
TransStart Green qPCR SuperMix UDG
TransStart
Top Green qPCR SuperMix
TransScript
Green Two-Step qRT-PCR SuperMix
TransScript
II Green Two-Step qRT-PCR SuperMix
TransScript
Green One-Step qRT-PCR SuperMix
TransScript II Green One-Step qRT-PCR SuperMix
TransStart
Probe qPCR SuperMix
TransScript Probe One-Step qRT-PCR SuperMix TransScript
II Probe One-Step qRT-PCR SuperMix
High Purity dNTPs
High Purity dNTPs
Chapter 1 PCR RT-PCR qPCR and qRT-PCR
059
060
062
063
064
066
068
070
072
073
TransScript
All-in-One First-Strand cDNA Synthesis
SuperMix for PCR
TransScript
All-in-One First-Strand cDNA Synthesis
SuperMix for qPCR
TransScriptII First-Strand cDNA Synthesis SuperMix
TransScript
II One-Step gDNA Removal and cDNA
Synthesis SuperMix
TransScript
II All-in-One First-Strand cDNA Synthesis
SuperMix for PCR
TransScript II All-in-One First-Strand cDNA SynthesisSuperMix for qPCR
TransScript
Two-Step RT-PCR SuperMix
TransScriptII Two-Step RT-PCR SuperMix
EasyScript One-Step RT-PCR SuperMix
TransScript
One-Step RT-PCR SuperMix
TransScript II One-Step RT-PCR SuperMix
Ribonuclease Inhibitor
046
047
048
049
050
051
052
053
054
055
056
058
075
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PCR EnzymesPCR enzymes are puried from E.coli strains carrying genes for specic DNA polymerase. The choice of DNA polymerase
for PCR application is highly dependent on the characteristics of the system as well as the desired results. The following is
PCR Enzyme Selection Guide.
PCR Enzyme Selection Guide
Application
PCR Enzyme Properties
GC/AT-rich
complex template
TransTaq HiFi
TransStart seriesGC Enhancer
TransTaq HiFi
TransStart series
TransDirect series
routine PCR high fidelity PCR
TransTaq series
TransStart TaqTransPfu series
Easy Taq
TransFast Taq
high specificity PCR
TransStart series
long PCR
PCR inhibitor
enriched template
PCR Enzyme TransFastTaq EasyTaq TransTaq-T TransTaqHiFi TransStartTaq TransStartTopTaq
Amplication Efciency TransFast
Taq =EasyTaq
TransTaq-T TransTaqHiFi TransStartTaq TransStartTopTaq
Specicity TransFast
Taq =EasyTaq
TransTaq
-T TransTaq
HiFi TransStart
Taq TransStart
TopTaq
Fidelity(vs EasyTaq) 1 1 18 18 18 18
Extension Rate 6 kb/min 1-2 kb/min 1-2 kb/min 1-2 kb/min 1-2 kb/min 1-2 kb/min
Hot start - - + + + +
3-5 exo + + + + + +
Product Size (human
genomic DNA as
template
4 kb 4 kb 8 kb 12 kb 12 kb 12 kb
Specication
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Applications
PCR Enzyme Applications
TransFastTaqDNA Polymerase routine PCR; fast PCR
EasyTaq DNA Polymerase routine PCR
TransTaq-T DNA Polymerase highly complex templates; TA cloning
TransTaqHiFi DNA Polymerase GC/AT; complex templates; long PCR; TA cloning
TransStartTaqDNA Polymerase qPCR; multiplex PCR; TA cloning
TransStartTopTaqDNA Polymerase qPCR; multiplex PCR; TA cloning
EasyPfuDNA Polymerase high delity PCR; blunt cloning; site-directed mutagenesis
TransStartFastPfuDNA Polymerase high delity PCR; fast PCR; robust PCR; blunt cloning; site-directed mutagenesis
TransStartFastPfuFlyDNA Polymerase high delity PCR; fast PCR; robust PCR; blunt cloning; site-directed mutagenesis
PCR Enzyme EasyPfu TransStart FastPfu TransStartFastPfu Fly
Amplication Efciency EasyPfu
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Description
TransFastTaq DNA polymerase is an engineered version ofTaq
DNA polymerase. The enzyme consists of a single polypeptide with a
molecular weight of approximately 94 kDa.
TransFastTaq DNA polymerase has 5-3 DNA polymerase activity
and 5-3 exonuclease activity. It lacks 3-5 exonuclease activity.
The extension rate is about 6 kb/min.
Template-independent A can be generated at the 3 end of the PCR
product. PCR products can be cloned in TA cloning vector.
Amplication of DNA fragment up to 4 kb.
Unit Denition
One unit ofTransFastTaqDNA Polymerase incorporates 10 nmol
of deoxyribonucleotide into acid-preceptible material in 30 minutes at
74oC.
Appl ications
Routine PCR.
Fast PCR.
Colony PCR.
Quality Contro l
TransFastTaqDNA polymerase has passed the following quality
control assays: functional absence of double-and single-stranded
endonuclease activity; >99% homogeneous measured by SDS-PAGE
gel electrophoresis. Each batch ofTransFastTaq DNA Polymerase
is assayed for amplication efciency from as little as 10 ng of human
genomic DNA.
Component Volume Final ConcentrationTemplate DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
10TransFastTaq Buffer 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
TransFastTaq DNA Polymerase 0.5 -1 l 2.5-5 units
ddH2O Variable -
Total volume 50 l -
Reaction Components
PROTOCOL
TransFastTaqDNA Polymerase
dNTPs
(2.5 mM)
AP101-11 500 units
AP101-12 6500 units
Concentration
5 units/l
Components
TransFastTaqDNA Polymerase
10TransFast Taq Buffer (200 mM Tris-HCl
pH 8.4; 100 mM KCl; 100 mM (NH4)2SO4;
20 mM MgSO4; others)
6DNA Loading Buffer
Storage
at -20oC for two years
dNTP-freeAP101-01 500 units
AP101-02 6500 units
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Description
EasyTaq DNA polymerase is purifed from E. coli expressing a cloned
Thermus aquaticus DNA polymerase. The enzyme consists of a
single polypeptide with a molecular weight of approximately 94 kDa.
EasyTaq
DNA polymerase has 5-3 DNA polymerase activity and 5-3
exonuclease activity. lt lacks 3-5 exonuclease activity. EasyTaq DNA
polymerase is suitable for routine amplication. PCR products are not
suitable for PAGE.
The extension rate is about 1-2 kb/min.
Template-independent A can be generated at the 3 end of the PCR
product. PCR products can be cloned in TA cloning vector.
Amplication of DNA fragments up to 4 kb.
Appl ication
Routine PCR.
Unit Denition
One unit ofEasyTaqDNA Polymerase incorporates 10 nmol of into acid-
preceptible material in 30 minutes at 74oC.
EasyTaqDN A Polymerase
Concentration
5 units/l
Components
EasyTaq - DNA Polymerase
10EasyTaq Buffer (200mM Tris-HCl
pH8.3; 200 mM KCl; 100 mM (NH4)2SO4;
20 mM MgSO4; others)
6DNA Loading Buffer
Storage
at -20oC for two year
Amplification Rate
0 50 100 150 200 250 300
1 Kb
2 Kb
3 Kb
4 Kb
min
EasyTaq
TransFast Taq
EasyTaq and TransFast Taq
Comparison Between
M ME F E F E F E F E F E F E F E F
M1Kb Plus DNA Ladder
EEasyTaq DNA Polymerase
FTransFast Taq DNA Polymerase
94oC 3 min
94oC 5 sec
50-60oC 15 sec 30-35 cycles
72o
C x sec72oC 5-10 min
0-2 kb
2-3 kb
>3 kb
10 sec/kb
20 sec/kb
30 sec/kb
dNTP-free
AP111-01 500 units
AP111-02
AP111-03
6500 units
42500 units
dNTPs
(2.5 mM)
AP111-11 500 units
AP111-12
AP111-13
6500 units
42500 units
Thermal cycling conditions
Target Extension time
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EasyTaqDNA Polymerase for PAGE
Concentration
5 units/l
94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 1-2 kb/min
72oC 5-10 min
M: 1Kb Plus DNA Ladder
1:CCRD 0.5 kb; 2: BDNF 0.8 kb;
3:Rhod 1.2 kb; 4: Rhod 2 kb;
5:Rhod 4.17 kb.
50 ng of Human Genomic DNA as templates
Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
10EasyTaq
Buffer 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
EasyTaq DNA Polymerase 0.5-1 l 2.5-5 units
ddH2O Variable -
Total volume 50 l -
Notes
MgSO4 is included in the 10PCR Buffer
at a nal concentration of 2.0 mM, which is
sufcient for most amplications. If needed,
suggested to use the 100 mM MgSO4
stock to prepare a titration from 2.0 mM to
4.0 mM (final concentration) in 0.25 mM
increments.
2.5 units ofEasyTaq DNA Polymerase is
enough for most target amplifications. For
some amplifications, up to 5 units/reaction
EasyTaq
DNA Polymerase can be used.
Reaction Components
PROTCOL
Thermal cycling conditions
Quality Control
EasyTaq DNA polymerase has passed the following quality control
assays: functional absence of double- and single-stranded
endonuclease activity; >99% homogeneous measured by SDS-PAGE
gel electrophoresis. EasyTaq DNA Polymerase is assayed foramplication efciency from as little as 10 ng of human genomic DNA.
dNTP-free
(2.5 mM)
AP112-01 2500 units
AP112-02 42500 units
dNTPs
(2.5 mM)
AP112-11 2500 units
AP112-12 42500 units
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Components
EasyTaqDNA Polymerase for PAGE
10EasyTaq Buffer for PAGE (200mM
Tris-HCl pH8.3; 200 mM KCl; 100 mM
(NH4)2SO4; 20 mM MgSO4; others)
6DNA Loading Buffer
Storage
at -20oC for two years
Description
EasyTaq DNA polymerase for PAGE is purifed from E. coli expressing
a cloned Thermus aquaticus DNA polymerase. The enzyme consists
of a single polypeptide with a molecular weight of approximately 94
kDa. EasyTaq
DNA polymerase for PAGE has 5-3 DNA polymeraseactivity and 5-3 exonuclease activity. lt lacks 3-5 exonuclease activity.
EasyTaq
DNA polymerase is suitable for routine amplication.
The extension rate is about 1-2 kb/min.
Template-independent A can be generated at the 3 end of the PCR
product. PCR products can be cloned in TA cloning vector.
Amplication of DNA fragments up to 4 kb.
Appl ication
Short fragment PCR.
Unit Denition
One unit ofEasyTaq
DNA Polymerase for PAGE incorporates 10 nmolof deoxyribonucleotide into acid-preceptible material in 30 minutes at
74oC.
94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 1-2 kb/min
72oC 5-10 min
Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
10EasyTaqBufferfor PAGE 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
EasyTaqDNA Polymerase for PAGE 0.5-1 l 2.5-5 units
ddH2O Variable -
Total volume 50 l -
Reaction Components
PROTOCOL
Thermal cycling conditions
Quality Contro l
EasyTaq DNA polymerase for PAGE has passed the following quality
control assays: functional absence of double- and single-stranded
endonuclease activity; >99% homogeneous measured by SDS-PAGE
gel electrophoresis. EasyTaq DNA Polymerase for PAGE is assayed
for amplication efciency from as little as 10 ng of human genomic
DNA.
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Description
TransTaq-T DNA Polymerase is an engineered version ofTaq DNA
polymerase. TransTaq-T DNA Polymerase has end transferase activity
and exonuclease activity. TransTaq-T DNA Polymerase combines the
amplication efciency ofTaq DNA polymerase with the delity ofPfu
DNA polymerase.
dNTP-free
AP122-01 250 units
AP122-02 500 units
AP122-03 6500 units
dNTPs(2.5 mM)
AP122-11 250 units
AP122-12 500 units
AP122-13 6500 units
TransTaq-T DNA Polymerase
Concentration
5 units/l
Components
TransTaq-T DNA Polymerase
10TransTaq-TBuffer (200 mM
Unit Denition
One unit ofTransTaq-T DNA Polymerase incorporates 10 nmol of
deoxyribonucleotide into acid-preceptible material in 30 minutes at 74oC.
Quality Control
TransTaq
-T DNA polymerase has passed the following qualitycontrol assays: functional absence of double-and single-stranded
endonuclease activity; >99% homogeneous measured by SDS-PAGE
gel electrophoresis. Each batch ofTransTaq-T DNA Polymerase is
assayed for amplication efciency from as little as 10 ng of human
genomic DNA.
Component Volume Final Concentration
Template DNA Variable as requiredForward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
10TransTaq-TBuffer 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
TransTaq-T DNA Polymerase 0.5-1 l 2.5-5 units
ddH2O Variable -
Total volume 50 l -
Notes
MgSO4 is included in the 10PCR Buffer
at a nal concentration of 2.0 mM, which
is sufcient for most amplications. If
needed, suggested to use the 100 mM
MgSO4 stock to prepare a titration from 2.0
mM to 4.0 mM (nal concentration) in 0.25
mM increments.
2.5 units ofTransTaq-T DNA polymerase
is enough for most target amplications.
For some amplications, up to 5 units/
reaction TransTaq -T DNA polymerase
can be used.
Reaction Components
PROTOCOL
Fidelity is 18 times higher than EasyTaqDNA Polymerase.
Template-independent A can be generated at the 3 end of the PCR
product. PCR products can be cloned in TA cloning vector. The extension rate is about 1-2 kb/min.
Amplication of DNA fragment up to 8 kb.
Appl ications
Complex PCR.
TA cloning.
Tris-HCl pH9.0; 100 mM KCl; 100 mM
(NH4)2SO4; 20 mM MgSO4; others)
6DNA Loading Buffer
Storage
at -20oC for two years
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Description
TransTaqDNA Polymerase High Fidelity (TransTaqHiFi DNA Polymerase)
isTransTaq
-T DNA Polymerase mixed with an 3-5exonuclease. TransTaq
HiFiDNA polymerase provides higher specicity and amplication efciency
thanTransTaq-T DNA Polymerase. The delity ofTransTaqHiFi DNA
polymerase is similar to EasyPfu DNA Polymerase.
Fidelity is 18 times higher than EasyTaq DNA Polymerase.
The extension rate is about 1-2 kb/min.
Template-independent A can be generated at the 3 end of the PCR
product. PCR products can be cloned in TA cloning vector
Amplication of DNA fragment up to 12 kb.
Appl ications
High delity.
High yield.
TransTaqHiFi Buffer I for genomic DNA, TransTaqHiFi Buffer II for
DNA, cDNA, Plasmid DNA amplication.
Unit Denition
One unit ofTransTaq HiFi DNA Polymerase incorporates 10 nmol of
deoxyribonucleotide into acid-preceptible material in 30 minutes at
74oC.
TransTaqDNA Polymerase High FidelityHiFi
Concentration
5 units/l
Components
TransTaqHiFi DNA Polymerase
10TransTaq HiFi Buffer I, II (200
mM Tris-HCl pH 9.0; 100 mM KCl; 100
mM (NH4)2SO4; 20 mM MgSO4; 10%
Glycerol,others)
6DNA Loading Buffer
GC Enhancer
Storage
at -20oC for two years
dNTPs
(2.5 mM)
AP131-11 250 units
AP131-12 500 units
AP131-13 6500 units
dNTP-free
AP131-01 250 units
AP131-02 500 units
AP131-03 6500 units
94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72
o
C 1-2 kb/min72oC 5-10 min
1 2 3 4 5 6 7M
M: 1Kb Plus DNA Ladder
1:BDNF 0.8 kb; 2: Rhod 1.2 kb;
3:-globin 1.3 kb; 4: Rhod 2.0 kb ;
5:-globin 3.0 kb; 6: Rhod 4.17 kb;
7: Factor IX 7.5 kb
50 ng of Human Genomic DNA as templates
M
Thermal cycling conditions
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Notes MgSO4 is included in the 10PCR Buffer
at a nal concentration of 2.0 mM, which
is sufcient for most amplications. If
needed, suggested to use the 100 mM
MgSO4 stock to prepare a titration from 2.0
mM to 4.0 mM (nal concentration) in 0.25
mM increments.
2.5 units ofTransTaqHiFi DNA
polymerase is enough for most target
amplications. For some amplications,
up to 5 unit/reaction TransTaqHiFi DNA
polymerase can be used.
PROTOCOL
Reaction Components
Quality Control
TransTaqHiFi DNA polymerase has passed the following quality
control assays: functional absence of double- and single-stranded
endonuclease activity; >99% homogeneous measured by SDS-PAGE
gel electrophoresis. Each batch ofTransTaq
HiFi DNA Polymeraseis assayed for amplication efciency from as little as 10 ng of human
genomic DNA.
Components Volume Final Concentration
Template DNA variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
10TransTaqHiFi Buffer I/II 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
TransTaq
HiFi DNA Polymerase 0.5-1 l 2.5-5 units
ddH2O to 50 l Not applicable
M1: 1Kb Plus DNA Ladder
M2:Trans15K DNA Marker
Lane 1: REPA 1.8 kb; Lane 2: NCBP 2.5 kb;
Lane 3: HDP 3.5 kb; Lane 4: VIN 4.6 kb;
Lane 5: Pol 6.8 kb; Lane 6: APC 8.5 kb;
Lane 7: Dynein 12.3 kb
100 ng Human total RNA as templates
TransTaq
HiFi Buffer II
TransTaq
HiFi Buffer I
M1: 1Kb Plus DNA LadderM2:Trans15K DNA MarkerLane 1: BDNF 0.8 kb;
Lane 2: Rhod 1.2 kb
Lane 3: Rhod 2.0 kb;
Lane 4: -globin 3.0 kb
Lane 5: Rhod 4.17 kb;
Lane 6: Factor IX 7.5 kb
Lane 7: Serum albumin 12.4 kb
50 ng Human Genomic DNA as templates
Thermal cycling conditions94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 1-2 kb/min
72oC 5-10 min
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Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
10TransTaqHiFi Buffer I/II 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
TransTaqHiFi DNA Polymerase 0.5-1 l 2.5-5 units
ddH2O Variable -
Total volume 50 l -
Notes MgSO4 is included in the 10PCR Buffer
at a nal concentration of 2.0 mM, which
is sufcient for most amplications. If
needed, suggested to use the 100 mM
MgSO4 stock to prepare a titration from 2.0
mM to 4.0 mM (nal concentration) in 0.25
mM increments.
2.5 units ofTransTaqHiFi DNA
polymerase is enough for most target
amplications. For some amplications,
up to 5 unit/reaction TransTaqHiFi DNA
polymerase can be used.
PROTOCOL
Reaction Components
Thermal cycling conditions94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 1-2 kb/min
72oC 5-10 min
TransStartTaqDNA Polymerase
Description
TransStartTaq DNA Polymerase is a hot start Taq DNA polymerase.
At room temperature, one binding protein binds to double-stranded
DNA template and another binding protein binds to primers. These
unique formulations effectively neutralize the DNA polymerase activity
at room temperature. The extension rate for TransStartTaq DNA
Polymerase is 1-2 kb/min. Template-independent A can be generated
at the 3 end of the PCR product. PCR products can be cloned in TA
cloning vector.
Appl ications Complex template.
GC/AT-rich template.
Multiplex PCR.
Unit Denition
One unit ofTransStartTaq DNA Polymerase incorporates 10 nmol of
deoxyribonucleotide into acid-insoluble material in 30 min at 74oC.
Quality Control
TransStartTaq DNA polymerase has passed the following quality
control assays: functional absence of double- and single-stranded
endonuclease activity; >99% homogeneous measured by SDS-PAGE
gel electrophoresis. Each batch of TransStartTaq DNA Polymerase
is assayed for amplication efciency from as little as 10 ng of human
genomic DNA.
Concentration
2.5 units/l
Components
TransStartTaq DNA Polymerase
10TransStart TaqBuffer (500 mM Tris-
HCl pH 9.0; 200 mM (NH4)2SO4; 20 mM
MgSO4; 10% Glycerol; others)
6DNA Loading Buffer GC Enhancer
Storage
at -20oC for two years
dNTP-free
AP141-01 250 units
AP141-02 500 units
AP141-03 6500 units
dNTPs
(2.5 mM)
AP141-11 250 unitsAP141-12 500 units
AP141-13 6500 units
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Notes
MgSO4 is included in the 10PCR Buffer
at a nal concentration of 2.0 mM, which is
sufcient for most amplications. If needed
suggested to use the 100 mM MgSO4
stock to prepare a titration from 2.0 mM to
4.0 mM (final concentration) in 0.25 mM
increments.
Choose GC Enhancer for the amplication
of GC/AT rich and complex template.
PROTOCOL
Reaction Components
TransStartHot Start ( Double Blocking )
TransStartdouble blocking technique blocks primer-dimer formation and polymerase activity at room temperature by blocking primers and DNA
templates at room temperature. Blocking proteins are released during the initial denaturation. This double blocking method has higher efciency
than antibody based, or chemical modied hot start PCR.
Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
10TransStartTaq Buffer 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
TransStartTaq DNA Polymerase 0.5-1 l 1.25-2.5 units
ddH2O Variable -
Total volume 50 l -
M1: 1Kb Plus DNA Ladder
M2:Trans15K DNA Marker
1: REPA 1.8 kb 2: NCBP 2.5 kb
3: HDP 3.5 kb 4: VIN 4.6 kb
5: Pol 6.8 kb 6: APC 8.5 kb
7: Dynein 12.3 kb
Human cDNA as templates
TransTaq
HiFi Buffer IITransTaq HiFi Buffer I
M1: 1Kb Plus DNA Ladder
M2:Trans15K DNA Marker
1: BDNF 0.8 kb 2: Rhod 1.2 kb
3: Rhod 2.0 kb 4: -globin 3.0 kb
5: Rhod 4.17 kb 6: Factor IX 7.5 kb
7: Serum albumin 12.4 kb
50 ng of Human Genomic DNA as
templates
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Thermal cycling conditions94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 1-2 kb/min
72oC 10 minM1 1Kb Plus DNA LadderM2 Trans15K DNA Marker
1 Numb 0.3 kb 2 CCRD 0.5 kb 3 BDNF 0.8 kb 4 Rhod 1.2 kb5 Rhod 2.0 kb 6 -globin 3.0 kb 7 Rhod 4.17 kb 8 Factor IX 7.5 kb9 Serum albumin 12.4 kb50 ng of Human Genomic DNA as templates
TransStartTopTaqDNA Polymerase
Description
TransStart
TopTaq DNA Polymerase is an engineered version ofTaq
DNA Polymerase combined with TransStartTechnique. TransStartdouble
blocking technique prevents primer-dimer formation and polymerase
activity at room temperature by blocking the primers and DNA templates
at room temperature. Blocking proteins are released from primers and
templates during the initial denaturation. This double blocking method has
higher efciency than antibody based, or chemical modied hot start PCR.
Compare with the TransStart
Taq DNA Polymerase, the TransStart
TopTaq DNA Polymerase has higher amplication efciency and
sensitivity. Fidelity is 18 times higher than EasyTat
DNA Polymerase.
The specicity is higher than antibody based chemical or modied hot
start DNA polymerases.
Reduced nonspecic amplication and primer dimers.
Different from Taq antibody, no risk of contamination from mammal
DNA.
Different from chemical modication , heating step no longer needed.
Amplication of DNA fragments up to 12 kb.
Appl ications Complex PCR.
Multiplex PCR.
High delity and specicity PCR.
High yield.Unit Denition
One unit ofTransStartTopTaq DNA Polymerase incorporates 10 nmol
of deoxyribonucleotide into acid-insoluble material in 30 min at 74oC.
Quality Control
TransStart
TopTaq DNA polymerase has passed the following quality
control assays: functional absence of double- and single-stranded
endonuclease activity; >99 % homogeneous measured by SDS-PAGE
gel electrophoresis. Each lot ofTransStart
TopTaq DNA Polymerase
is assayed for amplication efciency from as little as 10 ng of human
genomic DNA.
Concentration
2.5 units/l
Components
TransStartTopTaq DNA Polymerase
10TransStartTopTaqBuffer (500 mM
Tris-HCl (pH 9.0); 200 mM (NH4)2 SO4;
20 mM MgSO4; others)
6DNA Loading Buffer
GC Enhancer
Storage
at -20oC for two years
dNTP-free
AP151-01 250 units
AP151-02 500 units
AP151-03 6500 units
dNTPs
(2.5 mM)
AP151-11 250 units
AP151-12 500 units
AP151-13 6500 units
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Notes
MgSO4 is included in the 10PCR Buffer
at a final concentration of 2.0 mM, which issufficient for most targets amplificaiton. For
some targets, adjust the Mg2+ from 2.0 mM to
4.0 mM (nal concentration) to ensure a good
amplication.
Reaction Components
Thermal cycling conditions
94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 1-2 kb/min
72oC 5-10 min
M: Trans2K
Plus II DNA Marker
Lane 1: Rhod 1.2 kb Lane 2: Rhod 2.0 kb Lane 3: -globin 3.0 kb
Lane 4: -globin 4.1 kb Lane 5: Rhod 4.17 kb Lane 6: -globin 6.1 kb
Lane 7: Factor IX 7.5 kb Lane 8: IGF2R 8.9 kb Lane 9: Serum albumin 12.4 kb
1 2 3 4 5 6 7 8 9M
M: Trans2K
Plus DNA Marker
Lane 1: DMD1 0.3 kb Lane 2: Numb 0.3 kb
Lane 3: P53 0.5 kb Lane 4: P1P2 1.2 kb
Lane A: A company hot start polymerase
Lane T: TransStartTopTaq DNA Polymerase
1 2 3 4
M A T A T A T A T
50 ng Human Genomic DNA as templates
Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
10TransStartTopTaq Buffer 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
TransStartTopTaq DNA Polymerase 0.5-1 l 1.25-2.5 units
ddH2O Variable -
Total volume 50 l -
PROTOCOL
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EasyPfuDNA Polymerase
Description
EasyPfuDNA Polymerase is an engineered version ofpfu DNA Polymerase
with enhanced yield and higher delity. EasyPfu polymerase possesses
a proofreading 3'-5' exonuclease activity. Its delity is 18 times higher
than EasyTaq DNA Polymerase. EasyPfu DNA Polymerase is suitable
for blunt-end cloning and site-directed mutagenesis. Amplification of
fragment up to 6 kb.
Appl ications High delity.
Blunt-end cloning.
Site-directed mutagensis.
Unit Denition
One unit ofEasyPfuDNA P olymerase incorporates 10 nmol of
deoxyribonucleotide into acid-insoluble material in 30 min at 74oC.
Quality Control
EasyPfu polymerase has passed the following quality control assays:
functional absence of double- and single-stranded endonuclease
activity, >99% homogeneous measured by SDS-PAGE gel
electrophoresis. Each batch ofEasyPfu DNA Polymerase is assayed for
amplication efciency from as little as 10 ng of human genomic DNA.
dNTP-free
AP211-01 250 units
AP211-02 500 units
AP211-03 6500 units
dNTPs
(2.5 mM)
AP211-11 250 units
AP211-12 500 units
AP211-13 6500 units
Concentration
2.5 units/l
Components
EasyPfuDNA Polymerase
10EasyPfuBuffer(200 mM Tris-HCl
pH 8.8; 100 mM (NH4)2SO4; 100 mM
KCl; 20 mM MgSO4; others)
50 mM MgSO4
6DNA Loading Buffer
Storage
at -20oC for two years
PROTOCOL
Reaction Components
Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
10EasyPfu Buffer 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
EasyPfu DNA Polymerase 1 l 2.5 units
ddH2O Variable -
Total volume 50 l -
Thermal cycling conditions
94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 0.5 kb/min
72oC 5-10 min
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M: 1Kb Plus DNA Ladder
1:CCRD 0.5 kb; 2: BDNF 0.8 kb; 3: Rhod 1.2 kb; 4: -globin 1.3 kb;
5: Rhod 2 kb; 6: -globin 3 kb; 7: Rhod 4.17 kb; 8: -globin 4.1 kb
50 ng of Human Genomic DNA as templates
1 2 3 4 5 6 7 8M M
TransStart
FastPfuDNA Polymerase
Description
TransStartFastPfu DNA Polymerase is a hot start, high fidelity,
and high processivity DNA polymerase. The fidility of TransStart
FastPfu DNA polymerase is 54-fold higher than TaqDNA polymerase.TransStart
FastPfu DNA polymerase has extension rate up to 4 kb/
min. PCR product amplied by TransStartFastPfu DNA polymerase is
blunt end and can be cloned into blunt cloning vector.
Appl ications
High delity PCR.
High yield and robust PCR.
Blunt end cloning.
Site-directed mutagenesis.
Unit Denition
One unit of TransStart FastPfu DNA Polymerase incorporates 10
nmol of deoxyribonucleotide into acid-insoluble material in 30 min
at 74oC.
Quality Control
TransStartFastPfu DNA Polymerase has passed the following quality
control assays: functional absence of double- and single-stranded
endonuclease activity; >99% homogeneous measured by SDS-PAGE
gel electrophoresis. Each lot ofTransStartFastPfu DNA Polymerase
is assayed for amplication efciency from as little as 10 ng of human
genomic DNA.
dNTP-free
AP221-01 250 units
AP221-02 500 units
AP221-03 6500 units
dNTPs
(2.5 mM)
AP221-11 250 units
AP221-12 500 units
AP221-13 6500 units
Concentration
2.5 units/l
Components
TransStartFastPfu DNA Polymerase
5TransStartFastPfu Buffer (100 mM
Tris-SO4 pH 9.2; 50 mM (NH4)2SO4; 200
mM KCl; 10 mM MgSO4; 10% Glycerol;
others)
50 mM MgSO4
6DNA Loading Buffer
PCR Stimulant
Storage
at -20oC for two years
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PROTOCOL
Suggested conditions
50 lreaction volume
Parameter Targets10 kb Targets10 kb cDNA Targets
Template100 ng Genomic DNA 200-500 ng Genomic DNA 1-2 l cDNA from RT reaction
5-30 ng Plasmid DNA 5-30 ng Plasmid DNA (50-500 ng starting RNA template)
MgSO4 Add 1-2 l of 50 mM MgSO4 to a nal concentration of 3-4 mM for target larger than 5 kb
Thermal cycling conditions
Number of cycles Temperature Plasmid or Genomic DNA cDNA
1 95oC 2 min 1 min
Plasmid or Genomic DNA: 30-35
cDNA: 40
95oC 20 sec 20 sec
Tm-5oC 20 sec 20 sec
72oC15 sec for targets1 kb
15-30 sec per kb for targets >1 kb
30 sec for targets1 kb
30 sec per kb for targets>1 kb
1 72oC 5 min 5 min
Reaction Components
Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
5TransStartFastPfu Buffer 10 l 1
2.5 mM dNTPs 5 l 0.25 mM
TransStartFastPfu DNA Polymerase 1 l 2.5 units
ddH2O Variable -
Total volume 50 l -
cDNA amplication with TransStartFastPfu DNA Polymerase
1 2 3 4 5M1 M2
M1 1Kb Plus DNA Ladder
M2 Trans15K DNA Marker
1 ACTR 3.5 kb;
2 VIN 4.6 kb;
3 Pol 6.8 kb;
4 APC 8.5 kb;
5 Dynein 12.3 kb;
2 hrs 14 min
2 hrs 34 min
3 hrs 09 min
3 hrs 41 min
4 hrs 48 min
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M: Trans15K DNA Marker
1:UDG 7.0 kb;
2:LN 10.0 kb;
3:Fang 14.7 kb;
Genomic DNA amplication with TransStartFastPfu DNA Polymerase
1 2 3 4 5M1 M2
M1 1Kb Plus DNA Ladder
M2 Trans15K DNA Marker
1 Rhod 2.0 kb;
2 -globin 3.0 kb;
3 Rhod 4.17 kb;
4 Factor IX 7.5 kb;
5 Serum albumin 12.4 kb;
1 hrs 19 min
1 hrs 27 min
1 hrs 29 min
3 hrs 25 min
4 hrs 48 min
1 2 3M M
Plasmid DNAamplication with TransStartFastPfu DNA Polymerase
1 hrs 36 min
1 hrs 55 min
2 hrs 26 min
TransStartFastPfuFly DNA Polymerase
dNTP-free
AP231-01 250 units
AP231-02 500 units
AP231-03 6500 units
dNTPs
(2.5 mM)
AP231-11 250 unitsAP231-12 500 units
AP231-13 6500 units
Concentration
2.5 units/l
Components
TransStartFastPfu Fly DNA Polymerase
5TransStartFastPfu FlyBuffer (100
mM Tris-SO4 pH 9.2; 50 mM (NH4)2SO4;
200 mM KCl; 10 mM MgSO4; 10%
Glycerol; others)
Description
TransStartFastPfu Fly DNA Polymerase is a hot start, high delity, and
high processivity DNA polymerase. The dility of TransStartFastPfu
Fly DNA polymerase is 108-fold higher than TaqDNA polymerase.
TransStartFastPfu Fly DNA polymerase has extension rate up to 6 kb/
min. PCR product amplied by TransStartFastPfu FlyDNA polymerase
is blunt end and can be cloned into blunt cloning vector.
Appl ications
High delity PCR.
GC rich template and template that has secondary struture.
High yield and robust PCR.
Blunt end cloning.
Site-directed mutagenesis.
Long-segment PCR.
50 mM MgSO4
6DNA Loading Buffer
PCR Stimulant
Storage
at -20oC for two years
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Unit Denition
One unit of TransStart FastPfu Fly DNA Polymerase incorporates
10 nmol of deoxyribonucleotide into acid-insoluble material in 30
min at 74oC.
Quality Contro lTransStartFastPfu Fly DNA Polymerase has passed the following
quality control assays: functional absence of double- and single-
stranded endonuclease activity; >99% homogeneous measured by
SDS-PAGE gel electrophoresis. Each lot ofTransStartFastPfu DNA
Polymerase is assayed for amplication efciency from as little as 10
ng of human genomic DNA.
Suggested conditions
50 lreaction volume
Parameter Targets10 kb Targets10 kb cDNA Targets
Template100 ng Genomic DNA 200-500 ng Genomic DNA 1-2 l cDNA from RT reaction
5-30 ng Plasmid DNA 5-30 ng Plasmid DNA (50-500 ng starting RNA template)
MgSO4 Add 1-2 l of 50 mM MgSO4 to a nal concentration of 3-4 mM for target larger than 5 kb
Thermal cycling conditions
Number of cycles Temperature Plasmid or Genomic DNA cDNA
1 95oC 2 min 1 min
Plasmid or Genomic DNA: 30-35cDNA: 40
95oC 20 sec 20 sec
Tm-5
o
C 20 sec 20 sec
72oC15 sec for targets1 kb
15-30 sec per kb for targets >1 kb
30 sec for targets1 kb
30 sec per kb for targets>1 kb
1 72oC 5 min 5 min
Reaction Components
Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
5TransStartFastPfu FlyBuffer 10 l 1
2.5 mM dNTPs 5 l 0.25 mM
TransStartFastPfu Fly DNA Polymerase 1 l 2.5 units
ddH2O Variable -
Total volume 50 l -
1
18
54
108
1X
18X
108X
54X
EasyPfu TranStart
FastPfu
TranStart
FastPfuFlyEasyTaq
Fide
lity
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Description
GC Enhancer is used with DNA Polymerase, to optimize PCR of complex
templates or GC/AT-rich templates. The stock concentration is 10,
and the working concentration can be varied between 0.5- 5.
Appl ication
Amplication of complex templates and/or GC/AT-rich templates.
GC Enhancer
(50 l reaction mixture as example)
Storage
at -20oC for two years
AG101-01 200 l
GC Enhancers Volume (l) Final Concentration
2.5 0.5
5 1
10 2
15 3
20 4
25 5
10 kb
7 kb
4 kb
Size
PCR Time4 kb: Genomic DNA; 7 kb and 10 kb: Plasmid DNA
Total Reaction Time
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PCR Stimulant
DescriptionPCR Stimulant is used to optimize PCR of complex templates or GC/
AT-rich templates. It is especially suitable forPfu enzymes. The stock
concentration is 5, and the working concentration can be varied
between0.5- 2.5.
Appl ication
Amplication for complex templates and/or GC/AT-rich templates.
Storageat -20oC for two years
AG111-01 200 l
(50 l reaction volume as example)
PCR Stimulants Volume (l) Final Concentration
5 0.5
10 1
20 2
25 2.5
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PCR Enzyme vs PCR SuperMix
PCR SuperMix(+dye)
electrophoresis
PCR SuperMix(-dye)
cloning and sequencing
PCR SuperMix Selection Chart
optimize the PCR system
simple and quick
PCR Enzyme PCR SuperMix
purify
PCR SuperMixesPCR SuperMix is a ready-to-use mixture of DNA polymerase, salt, magnesium, dNTPs, and other components for efcient
PCR amplication. All you have to do is to add template, primers, and dH2O. If using PCR SuperMixes with dye, dye needs
to be removed before sequencing.
Appl ications
PCR SuperMix Application
2EasyTaqPCR SuperMix routine PCR
2TransTaq-T PCR SuperMix complex templates and TA cloning
2TransTaqHigh Fidelity(HiFi)
PCR SuperMix I
GC/AT-rich templates, complex templates
long fragment amplication, genomic DNA amplication (
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PROTOCOL
Total volume 50 l -
EasyTaq
PCR SuperMix (+dye) EasyTaq
PCR SuperMix (-dye)
M
M: 1Kb Plus DNA Ladder
1:CCRD 0.5 kb
2:BDNF 0.8 kb
3:Rhod 1.2 kb
4: Rhod 2 kb
5:Rhod 4.17 kb
50 ng of Human Genomic DNA as
templates
M M 1 2 3 4 51 2 3 4 5
Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
2EasyTaqPCR SuperMix 25 l 1
ddH2O Variable -
Notes
Completely thaw the contents in the tube
before each use.
Reaction Components
Thermal cycling conditions
94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 1-2 kb/min
72oC 5-10 min
Description
EasyTaq PCR SuperMix is a ready-to-use mixture ofEasyTaq
DNA Polymerase, dNTPs, and optimized buffer. This SuperMix
is provided at 2 concentration and used at 1 concentration
by adding template, primers and H2O. PCR products are not
suitable for PAGE.
The extension rate is about 1-2 kb/min. Template-independent A can be generated at the 3 end of the PCR
product. PCR products can be cloned in TA cloning vector
Amplication of DNA fragment up to 4 kb.
Appl ication
Routine PCR.
Mix(-dye)
AS111-01 1 ml
AS111-02 51 ml
AS111-03 151 ml
Mix(+dye)
AS111-11 1 ml
AS111-12 51 ml
AS111-13 151 ml
2EasyTaqPCR SuperMix
Storage
at -20oC for two years
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PROTOCOL
Component Volume Final Concentration
Template DNA
Variableas required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
2EasyTaqPCR SuperMix for PAGE 25 l 1
ddH2O Variable -
Total volume 50 l -
Notes
Completely thaw the contents in the tube
before each use.
Reaction Components
Thermal cycling conditions
94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 1-2 kb/min
72oC 5-10 min
Description
EasyTaq PCR SuperMix for PAGE is a ready-to-use mixture of
EasyTaq DNA Polymerase for PAGE, dNTP s, and optimized
buffer.This SuperMix for PAGE is provided at 2 concentration
and used at 1 concentration by adding template, primers and
H2O.
The extension rate is about 1-2 kb/min.
Template-independent A can be generated at the 3 end of the PCR
product. PCR products can be cloned in TA cloning vector
Amplication of DNA fragment up to 3 kb.
Appl ication
PCR of short fragments.
2EasyTaqPCR SuperMix for PAGE
Storage
at -20oC for two years
Mix (+dye)
AS112-11 1 ml
AS112-12 51 ml
AS112-13 151 ml
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Description
TransTaq-T PCR SuperMix is a ready-to-use mixture ofTransTaq-T
DNA Polymerase, dNTPs, and optimized buffer. This SuperMix is
provided at 2concentration and used at 1concentration by adding
template, primers and H2O.
Fidelity is 18 times higher than EasyTaq DNA Polymerase.
Template-independent A can be generated at the 3 end of the PCR
product. PCR products can be cloned in TA cloning vector.
The extension rate is about 1-2 kb/min.
Amplication of DNA fragment up to 8 kb.
Appl ications
Complex template.
TA cloning.
Mix (-dye)AS122-01 1 ml
AS122-02 51 ml
Mix (+dye)AS122-11 1 ml
AS122-12 51 ml
2TransTaq-T PCR SuperMix
Storage
at -20OC for two years
Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
2TransTaq-T PCR SuperMix 25 l 1
ddH2O Variable -
Thermal cycling conditions
94 2-5 min
94 30 sec
50-60 30 sec 30-35 cycles
72 1-2 kb/min
72 5-10 min
PROTOCOL
Reaction ComponentsNotes
Completely thaw the contents in the tube
before each use.
TransTaq-T DNA Polymerase TransTaq
-T PCR SuperMix
MMM 1 2 3 4 5 6 7M: 1Kb Plus DNA Ladder
1:BDNF 0.8 kb; 2: Rhod 1.2 kb;
3:-globin 1.3 kb; 4: Rhod 2.0 kb;
5: -globin 3.0 kb; 6: Rhod 4.17 kb;
7: Factor IX 7.5 kb
50 ng of Human Genomic DNA as templates
1 2 3 4 5 6 7
Total volume 50 l -
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Component Volume Final Concentration
Template DNA Variable as required
Forward Primer (10 M) 1-2 l 0.2-0.4 M each
Reverse Primer (10 M) 1-2 l 0.2-0.4 M each
2TransTaqHiFi PCR SuperMix 25 l 1
ddH2O Variable -
Reaction Components
PROTOCOL
DescriptionTransTaq
High Fidelity (HiFi) PCR SuperMix I or II is a ready-to-
use mixture ofTransTaq High Fidelity(HiFi) DNA polymerase (a
hot start DNA polymerase ), with a proofreading 3-5 exonuclease,
dNTPs, and optimized buffer. TransTaq High Fidelity (HiFi) PCR
SuperMix I is designed for the amplification of genomic DNA, and
TransTaqHigh Fidelity (HiFi) PCR SuperMix II is designed for the
amplication of DNA, cDNA, or plasmid DNA. The delity is 18 fold
higher than EasyTaq DNA polymerase. This SuperMix is provided at 2
concentration and can be used at 1concentration by adding template,
primers and H2O.
Fidelity is 18 times higher than EasyTaq DNA Polymerase. The extension rate is about 1-2 kb/min.
Template-independent A can be generated at the 3 end of the PCR
product. PCR products can be cloned in TA cloning vector.
Amplication of DNA fragment up to 12 kb.
Appl ications
High delity.
High yield.
Mix I(-dye)AS131-01 1 ml
AS131-02 51 ml
Mix II(-dye)AS131-21 1 ml
AS131-22 51 ml
2TransTaqHigh FidelityHiFiPCR SuperMix
Storage
at -20oC fortwo years
Notes
Completely thaw the contents in the tubebefore each use.
Thermal cycling conditions
94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 1-2 kb/min
72oC 5-10 min
Total volume 50 l -
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TransTaq
HiFi DNA Polymerase TransTaq
HiFi PCR S uperMix II
M1 1Kb Plus DNA Ladder
M2 Trans15K DNA Marker
1:REPA 1.8 kb; 2: NCBP 2.5 kb; 3: HDP 3.5 kb; 4: VIN 4.6 kb;
5:Pol 6.8 kb; 6:APC 8.5 kb; 7: Dynein 12.3 kb
Human cDNA as templates
M21 2 3 4 5 6 7 M2M1 M1 1 2 3 4 5 6 7
TransTaq
HiFi DNA Polymerase TransTaq
HiFi PCR SuperMix I
M1 1Kb Plus DNA Ladder
M2 Trans15K DNA Marker
1:BDNF 0.8 kb; 2:Rhod 1.2 kb; 3: Rhod 2.0 kb; 4: -globin 3.0 kb;
5:Rhod 4.17 kb; 6: Factor IX 7.5 kb; 7: Serum albumin 12.4 kb
50 ng of Human Genomic DNA as templates
M21 2 3 4 5 6 7 M2M1 M1 1 2 3 4 5 6 7
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Description
EasyPfuPCR SuperMix is a ready-to-use mixture ofEasyPfu DNA
polymerase , dNTPs, and optimized buffer. This SuperMix is provided
at 2concentration and used at 1concentration by adding template,
primers and H2O. PCR products can be cloned into pEASY-Blunt
cloning vector series.
Appl ications
High delity PCR amplication.
Blunt end cloning.
Site-directed mutagenesis.
Mix (-dye)AS211-01 1 ml
AS211-02 51 ml
2EasyPfuPCR SuperMix
Storage
at -20oC for two years
PROTOCOL
Thermal cycling conditions
94oC 2-5 min
94oC 30 sec
50-60oC 30 sec 30-35 cycles
72oC 0.5 kb/min
72oC 5-10 min
M: 1Kb Plus DNA Ladder
1:CCRD 0.5 kb; 2: BDNF 0.8 kb; 3: Rhod 1.2 kb; 4: -globin 1.3 kb;
5: Rhod 2 kb; 6: -globin 3 kb; 7: Rhod 4.17 kb; 8: -globin 4.1 kb
50 ng of Human Genomic DNA as templates
1 2 3 4 5 6 7 8 M1 2 3 4 5 6 7 8
EasyPfu DNA Polymerase 2xEasyPfu PCR SuperMix
MM
Notes
Completely thaw the contents in the tube
before each use.Component Volume Final Concentration
Template DNA Variable
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Description
TransStart
FastPfu PCR SuperMix is a ready-to-use mixture of
TransStartFastPfu DNA polymerase, dNTPs, and optimized buffer.
This SuperMix is provided at 2concentration and can be used at
1concentration by adding template, primers and H2O. PCR product
amplied by TransStartFastPfu DNA polymerase can be cloned into
pEASY-Blunt cloning vector series.
Extension rate up to 4 kb/min.
High delity, robust, and high speed amplication.
Appl ications
High delity amplication.
Site-directed mutagenesis.
Long or complex template amplication.
Mix (-dye)AS221-01 1 ml
AS221-02 51 ml
2TransStartFastPfu PCR SuperMix
Storage
at -20oC for two years
PROTOCOL
Reaction Components
Thermal cycling conditions94oC 2-5 min
94oC 20 sec
50-60oC 20 sec 30-35 cycles
72oC 2-4 kb/min
72oC 5-10 min
Notes
Completely thaw the contents in the tube
before each use.
Component Volume Final Concentration
Template DNA Variable
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Description
TransDirectTM Animal Tissue PCR Kit uses a propriety formulated
lysis reagent to release nucleic acids from a variety of animal
tissues(fresh,frozen). Unpurified DNA is used as template for PCR
using 2TransDirectTM PCR SuperMix which has extremely high
resistance to many PCR inhibitors found in animal tissues.
TransDirect
TM
Animal Tissue PCR Kit
Storage
at -20oC for two years
Kit Components
Amount of Starting Mater ial
Component AD201-01 AD201-02
AD1 Buffer 12 ml 55 ml
AD2 Preparation Buffer 3 ml 15 ml
AD3 Buffer 12 ml 512 ml
2TransDirectTM PCR SuperMix(+dye) 1 ml 51 ml
ddH2O 5 ml 25 ml
Material Amount
Mammalian Cell 1-5106 cell
Animal Tissues 10-30 mg
Mouse Tail 0.5-1 cm sections
Mouse Ear 0.5-0.7 cm disk
Saliva 10-30 l
Hair shaft 30 mg
Appl ications
Propriety formulated lysis reagent to release nucleic acids from avariety of animal tissues like mammalian cells, saliva, hair shaft,
animal .
Extremely high animal tissue inhibitor resistance 2TransDirectTM
PCR SuperMix for direct PCR from unpuried DNA.
Amplications up to 3 kb.
AD201-01 100 rxns20 l system
AD201-02 500 rxns20 l system
Direct PCR Application
TransDirectTM Animal Tissue PCR Kit mammalian cells, saliva, hair shaft, animal tissues
TransDirectTM Plant Tissue PCR Kit Non polysaccharides and polyphenols in plant tissue
TransDirectTM Blood PCR KitFresh and frozen blood sample stored in EDTA, heparin, or citric acid
Blood sample and dried blood without the anticoagulant
Direct PCRTransDirect PCR uses a propriety formulated lysis reagent to release nucleic acids from a variety of animal
tissues(fresh,frozen), plant tissues. Unpuried DNA is used as template for PCR using 2TransDirectTM PCR SuperMix which
has extremely high resistance to many PCR inhibitors found in animal tissues, plant tissues and blood.
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PROTOCOL
ProcedureNotes
Completely thaw the contents in the tube
before each use.
Components Volume Final Concentration
Tissue Extract 4 l as required
2TransDirectTM PCR SuperMix (+dye) 10 l 1
Forward Primer (10 M) 0.4-0.8 l 0.2-0.4 M each
Reverse Primer (10 M) 0.4-0.8 l 0.2-0.4 M each
ddH2O to 20 l Not applicable
A:Add 25 l of AD2 buffer into 100 l of the AD1 Buffer, mix thoroughlyby pipetting up and down (where substantial extracted sample, AD1
buffer to AD2 Buffer at ratio of 4:1, pre-mixed, and use within two
hours).
B:For different samples, the following processing methods:
Mammalian Cells
From collected cells, thoroughly remove the culture broth, add the
mixture of AD1 and AD2, mix thoroughly by pipetting up and down.
Animal Tissues
Cut using clean, sterile scissors or blade, add the mixture of AD1 and
AD2, mix thoroughly by pipetting up and down.
Saliva
Directly add saliva into the mixture of AD1 and AD2, mix thoroughly
by pipetting up and down.
Hair Shafts
Cut hair into pieces, add the mixture of AD1 and AD2, mix thoroughly
by pipetting up and down.
C:Incubate for 10 minutes at room temperature (for difficult to lyse
cells such as hair, 55oC incubated for 10 minutes, followed by 95oC
incubated for 10 minutes). After incubation, 3 minutes at 95 oC.
D: Adding 100 ul AD3 Buffer, mix directly as template for PCR, or placed
and stored at 4oC or -20oC.
Thermal cycling conditions94 5-10 min
94 30 sec
50-60 30 sec 35-40 cycles
72 1-2 kb/min
72 5-10 min
M
M:Trans2K
Plus II DNA Marker
Lane 1: Hair 0.8 kb
Lane 2: Saliva 0.8 kb
Lane 3: HeLa 0.8 kb
Lane 4: HeLa 2 kb
Lane 5: HeLa 3 kb
Lane 6: Mouse ear 0.86 kb
Lane 7: Mouse ear 1.8 kb
7 8 9 10 11 121 2 3 4 5 6 13 14
Lane 8: Mouse ear3 kb
Lane 9: Drosophila 0.42 kb
Lane 10: Drosophila 2 kb
Lane 11: Nematode 0.9 kb
Lane 12: Shrimp 1.1 kb
Lane 13: Crab 1 kb
Lane 14: Razor clan 0.56 kb
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Description
TransDirectTM Plant Tissue PCR Kit uses a propriety formulated
lysis reagent to release nucleic acids from a variety of plant tissues.
Unpurified DNA is used as template for PCR using 2TransDirectTM
PCR SuperMix which has extremely high resistance to many PCR
inhibitors found in plant tissues.
Appl ications
Propriety formulated lysis reagent to release nucleic acids from a
variety of plant tissues
Extremely high plant tissue inhibitor resistance 2TransDirectTM
PCR SuperMix for direct PCR from unpuried DNA or PCR from cell
culture without DNA extraction.Amplications up to 2 kb.
TransDirectTM Plant Tissue PCR Kit
Storage
at -20oC for two years
PROTOCOL
Notes
Completely thaw the contents in the tube
before each use.
AD301-01 100 rxns20 l system
AD301-02 500 rxns20 l system
Components AD301-01 AD301-02
PD1 Buffer 12 ml 55 ml
PD2 Buffer 12 ml 55 ml
2TransDirectTM PCR
SuperMix (+dye)1 ml 51 ml
ddH2O 5 ml 25 ml
Kit Components
Incubate the bottle for PD1 at 55oC until it becomes clear, if it is
viscous
A: Genomic DNA Extraction
1.Cut up 10-30 mg or 1cm2 plant tissue and add it to a tube containing
100 l of PD1 buffer, Vortex.
2.Incubate at 95C for 10 min.
3.Add 100 l of PD2 Buffer and vortex to mix. DNA is ready for PCR
or storage (Tissue Extract can be stored at 4oC for six months, or at
-20oC one year) .
B:PCR amplication
Components Volume Final Concentration
Tissue Extract 2-4 l as required
2TransDirectTM PCR SuperMix (+dye) 10 l 1
Forward Primer (10 M) 0.4-0.8 l 0.2-0.4 M each
Reverse Primer (10 M) 0.4-0.8 l 0.2-0.4 M each
ddH2O to 20 l Not applicable
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Description
Trans Blood PCR SuperMix is a ready-to-use mixture ofTransBD
Taq DNA Polymerase, dNTPs, and optimized buffer. This SuperMix is
provided at 2concentration and can be used at 1concentration by
adding blood, primers and H2O.
Appl ications
Fresh and frozen blood sample stored in EDTA, heparin, or
citric acid.
Blood sample and dried blood without the anticoagulant.
PCR from cell culture without DNA extraction.
Amplication upto 4 kb.
Mix (-dye)AD401-01 100 rxns20 l system
AD401-02 500 rxns20 l system
TransDirectTM Blood PCR Kit
Storage
at -20oC for two years
Thermal cycling conditions94 2-5 min
94 20 sec
50-60 20 sec 30-35 cycles
72 2-4 kb/min
72 5-10 min
M 7 8 9 10 11 121 2 3 4 5 6 13 14 15 16 17 18 19 20
M:Trans2K
Plus II DNA Marker
Lane 1: Corn 0.4 kb
Lane 2: Corn 0.8 kb
Lane 3: Corn 1.5 kb
Lane 4: Wheat 0.4 kb
Lane 5: Wheat 0.9 kb
Lane 6: Wheat 1.5 kb
Lane 7: Soybean 0.9 kb
Lane 8: Soybean1.5 kb
Lane 9: Arabidopsis 0.5 kb
Lane 10: Arabidopsis 0.9 kb
Lane 11: Arabidopsis 1.5 kb
Lane 12: Tobacco 0.5 kb
Lane 13: Tobacco 0.9 kb
Lane 14: Tobacoo 1.5 kb
Lane 15: Cotton 0.3 kb
Lane 16: Cotton 1 kb
Lane 17: Cotton 1.6 kb
Lane 18: Rice 0.3 kb
Lane 19: Rice0.9 kb
Lane 20: Rice 1.9 kb
Kit Components
Components AD401-01 AD401-02
2TransDirectTM PCR
SuperMix (+dye)1 ml 51 ml
ddH2O 5 ml 25 ml
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human frozen EDTA anticoagulated blood
human frozen heparin anticoagulated blood
human fresh blood sample without the anticoagulant
0.5 l of blood sample was used in 25 l PCR reaction.
Hela cells were used to amplify the Hdtgene (0.32 kb) in 25 l PCR reaction
M: Trans2K
DNA Marker
Lane 1: 104cells
Lane 2: 103cells
Lane 3: 102cells
Lane 4: 10 cells
Human oral cavity epithelial tissue wasused to amplify the Hdt gene (0.32 kb)
in 25 l reaction
M: Trans2K
DNA Marker
0.5 l of 1:40 diluted mouse heparin
anticoagulated blood was used in
25 l reaction .
M: Trans2K
DNA MarkerLane 1: Neo 0.25 kb
Lane 2: MAP 0.6 kb
0.5 l of diluted chicken sodium citrate
anticoagulated blood was used to amplifya 0.25 kb fragment in 25 l reaction
M: Trans2K
DNA MarkerLane 1: Blood
Lane 2: 1:10 dilution
Lane 3: 1:100 dilution
PROTOCOL
Reaction Components
Thermal cycling conditions
94oC 5 min
94oC 30 sec
50-60oC 30 sec 30-40 cycles
72o
C 1-2 kb/min72oC 5-10 min
Notes
Completely thaw the contents in the tube
before each use.
Components Volume Final Concentration
Template 0.2-0.8 l as required
Forward Primer (10 M) 0.4-0.8 l 0.2-0.4 M each
Reverse Primer (10 M) 0.4-0.8 l 0.2-0.4 M each
2TransDirectTMPCR SuperMix (+dye) 10 l 1
ddH2O to 20 l Not applicable
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RT-PCR
Transreverse transcriptase series include
EasyScript RT,TransScript RT andTransScript II RT.
Products Size ofproduction Sensitivity FidelityComplex or GC-rich
template
EasyScriptRT 8 kb
TransScriptRT 12 kb
TransScript
II RT 15 kb
EasyScriptFirst-Strand cDNA Synthesis SuperMix 8 kb
TransScriptFirst-Strand cDNA Synthesis SuperMix 12 kb
TransScriptOne-Step gDNA Removal and cDNA
Synthesis SuperMix12 kb
TransScriptAll-in-One First-Strand cDNA Synthesis
SuperMix for PCR12 kb
TransScriptAll-in-One First-Strand cDNA Synthesis
SuperMix for qPCR12 kb
TransScriptII First-Strand cDNA Synthesis SuperMix 15 kb
TransScript II One-Step gDNA Removal and cDNA
Synthesis SuperMix 15 kb
TransScript II All-in-One First-Strand cDNA Synthesis
SuperMix for PCR15 kb
TransScript II All-in-One First-Strand cDNA Synthesis
SuperMix for qPCR15 kb
TransScriptTwo-Step RT-PCR SuperMix 12 kb
TransScript II Two-Step RT-PCR SuperMix 15 kb
EasyScriptOne-Step RT-PCR SuperMix 4 kb
TransScriptOne-Step RT-PCR SuperMix 8 kb
TransScript II One-Step RT-PCR SuperMix 8 kb
TransScript
II RT
TransScript RT
EasyScript
RT
Trans reverse transcriptases
Site mutation tohave a higherthermostability
Site mutation conveyinghigher synthesis ability
Site mutation todestroy RNase Hactivity
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U A
NNNNGAAAAAAAAAAAAAAA+GTTTTTTTTTTTTC C
NNNNUAAAAAAAAAAAAAAAAATTTTTTTTTTTT
NNNNCAAAAAAAAAAAAAAAAGTTTTTTTTTTTT
NNNNGAAAAAAAAAAAAAAAA
CTTTTTTTTTTTT
UNNNNCAAAAAAAAAAAAAAAA+TTTTTTTTTTTT
G
UNNNNCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
G TTTTTTTTTTTT
Trans RT products (except one-step kits) use Anchored Oligo(dT) to increase cDNA yield and full length cDNA
products
Anchored oligo(dT) binds specic site.
Higher cDNA yield and more full length cDNA.
Poly(A) tail can be a few hundreds bp long and oligo(dT)
binds randomly.
Difcult to synthesize long poly(A) tail.
Lower yield and less full length cDNA.
Anchored Oligo(dT) PrimerTraditionalOligo(dT)12-18 Primer
Primer
RNA
2RT Reaction Mix
RT/RI Enzyme Mix
Tradtional RT vs Trans RT
Mix RNA and primers
Denature 5 min at 65oC,
place on ice
Add buffer RNase
Inhibitor dNTPs
M-MLV RT
Incubate at 42oC for
50 min , then 15 min at
70oC to inactive the
enzyme
Add RNA primers
buffer RNase Inhibitor
dNTPs Trans RT,
in one tube
Incubate at 42oC/50oC
for 30 min , then 5 minat 85oC to inactive the
enzyme
Add all components
without thermal
denaturation and
ice-bath
Traditional First-Strand cDNA Synthesis vs TransFirst
-Strand cDNA Synthesis SuperMix
TradtionalTraditional TransTrans
Primer
RNA
Buffer
dNTPs
RT Enzyme
RNase Inhibitor
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EasyScript Reverse Transcriptase[M-MLV,RNase H ]
AE101-02 10000 units
AE101-03 510000 units
Description
EasyScript Reverse Transcriptase is an engineered version of
M-MLV reverse transcriptase with reduced RNase H activity. The
enzyme is purified to near homogeneity from E.coli containing the
modied M-MLV RT gene.
Reduced RNase H activity to reduce RNA template degradation
during the rst-strand cDNA synthesis.
Anchored oligo(dT)18 for higher yield and more full length cDNA.
cDNA up to 8 kb.
Appl ication
First strand cDNA synthesis.
Multiple copy gene.
Unit Denition
One unit ofEasyScript is the amount of enzyme required to
incorporate 1 nmol of deoxyribonucleotide into acid precipitable
material in 10 minutes at 37oC using Poly(A)Oligo(dT) as template
primer.
Concentration
200 units/l
Components
EasyScriptRT
5ES RT Buffer (375 mM KCl;
15 mM MgCl2; 100 mM Tris-HCl pH 8.4)
Anchored Oligo(dT)18
Storage
at -20oC for one year
PROTOCOL
First Strand cDNA synthesis1 Reaction Components
Component Volume
Total RNA/mRNA Variable
Anchored Oligo(dT)18(0.5 g /l) 1 l
or Random Primer(N9) (0.1 g/l) 1 l
or GSP 2 pmol
10 mM dNTPs 1 l
5ES RT Buffer 4 l
Ribonuclease Inhibitor (50 units/l) 0.5 l
EasyScript
RT 1 l
RNase-free Water to 20 l
2 Incubation
For anchored oligo(dT)18primer or GSP, incubate at 42oC for 30 min.
For random primer, incubate at 25oC for 10 min, thenat 42oC for 30 min.
3 Incubate at 85oC for 5 min to inactive enzyme.
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Thermal cycling conditions
94oC 2-5 min
94
o
C 30 sec50-60oC 30 sec 35-40 cycles
72oC 1-2 kb/min
72oC 5-10 min
M 1Kb Plus DNA Ladder
1 GAPDH 0.5 kb 2 GAPDH 0.9 kb
3 REPA 1.8 kb 4 ACTR 3 kb
5 ACTR 3.5 kb 6 VIN 4.6 kb
7 TSC 5.3 kb 8 Pol 6.8 kb
Human cDNAas template
1 2 3 4 5 6 7 8M
NCBP Human DNA Polymerase
M M M1 2 3 4 5 6 7 1 2 3 4 5 6 7M 1Kb Plus DNA Ladder
Lanes 1-7 0 0.01 0.1 1 10 100 1000 pg
Human total RNA
M
Component Volume Final Concentration
cDNA Variable as requiredForward Primer (10 M) 1 l 0.2 M
Reverse Primer (10 M) 1 l 0.2 M
10TransTaqHiFi Buffer II (Mg2+
) 5 l 1
2.5 mM dNTPs 4 l 0.2 mM
TransTaqHiFi DNA Polymerase 0.5 l 2.5 units
ddH2O Variable -
Total volume 50 l -
RT-PCRReaction Components
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AT101-02 10000 units
AT101-03 510000 units
TransScriptReverse Transcriptase[M-MLV,RNase H ]
Concentration
200 units/l
Components
TransScript RT
5TS RT Buffer (250 mM KCl;
15 mM MgCl2; 100 mM Tris-HCl pH 8.4)
Anchored Oligo(dT)18
Storage
at -20oC for one year
NCBP
M M M
Human DNA Polymerase
1 2 3 4 5 6 7 1 2 3 4 5 6 7
M Trans15K DNA Marker
1 GAPDH 0.9 kb 2 REPA 1.8 kb
3 ACTR 3 kb 4 HDP 3.5 kb
5: VIN 4.6 kb 6 TSC 5.3 kb
7 Pol 6.8 kb 8 FIB 9.4 kb9 Dynein 12.3 kb
Human cDNA as template
M 1Kb Plus DNA Ladder
Lanes 1-7 0 0.01 0.1 1 10 100 1000 pg
Human total RNA
PROTOCOL
The condition for the rst strand cDNA synthesis is the same as for
EasyScript Reverse Transcriptase ( P39 ).
M1 2 3 4 5 6 7 8 9M
Description
TransScript Reverse Transcriptase is a recombinant M-MLV reverse
transcriptase with reduced RNase H activity.
Reduced RNase H activity to reduce RNA template degradation
during the rst-strand cDNA synthesis.
Anchored oligo(dT)18 for higher yield and more full length cDNA.
cDNA up to 12 kb.
Appl ication
First strand cDNA synthesis.
Multiple copy and low-copy genes.
Unit Denition
One unit ofTransScript is the amount of enzyme required to
incorporate 1 nmol of deoxyribonucleotide into acid-precipitable
material in 10 minutes at 37oC using Poly(A)Oligo(dT) as template
primer.
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First Strand cDNA synthesis
1 Reaction Components
2 Incubation
For anchored oligo(dT)20primer or GSP, incubate at 50oC for 30 min.
For random primer, incubate at 25oC for 5-10 min then at 50oC for 30
min.
For GC-rich or complex secondary structure RNA template, incubate
at 55oC for 30 min.
3 Incubate at 85oC for 5 min to inactive enzyme.
PROTOCOL
Component Volume
Total RNA/mRNA Variable
Anchored Oligo(dT)20(0.5 g /l) 1 l
or Random Primer(N9) (0.1 g/l) 1 l
or GSP 2 pmol
10 mM dNTPs 1 l
10TS II RT Buffer 2 l
Ribonuclease Inhibitor (50 units/l) 0.5 l
TransScript II RT 1 l
RNase-free Water to 20 l
TransScript II Reverse Transcriptase[M-MLV,RNase H ]
(High Temperature RT)
Description
TransScript II Reverse Transcriptase is a recombinant M-MLV
reverse transcriptase with reduced RNase H activity and increased
thermostability. The enzyme is active up to 55oC. It provides higher
specicity, higher yield and more full-length cDNA products.
Increased thermostability for more full-length cDNA products.
Reduced RNase H activity to reduce RNA template degradation
during the rst-strand cDNA synthesis.
Anchored oligo(dT)20 for higher yield and more full length cDNA.
cDNA up to 15 kb.Appl ication
First strand cDNA synthesis.
Multiple copy and low-copy genes.
GC rich or comlex secondary-structure RNA template.
Unit Denition
One unit ofTransScript II is the amount of enzyme required to
incorporate 1 nmol of deoxyribonucleotide into acid-precipitable
material in 10 minutes at 37oC using Poly(A)Oligo(dT) as template
primer .
Concentration
200 units/l
Components
TransScript II RT
10TS II RT Buffer (500 mM KCl; 30
mM MgCl2; 200 mM Tris-HCl pH 8.4)
Anchored Oligo(dT)20
Storageat -20oC for one year
AH101-02 10000 units
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M1 1Kb Plus DNA Ladder
M2 Trans15K DNA Marker
1: REPA 1.8 kb; 2: NCBP 2.5 kb;
3: HDP 3.5 kb; 4: VIN 4.6 kb;
5: Pol 6.8 kb; 6: APC 8.5 kb;
7: Dynein 12.3 kb; 8: FAL 15.1 kb
Human cDNA as templates
M21 2 3 4 5 6 7 8M1
The condition for the PCR is the same as for EasyScript Reverse
Transcriptase(P37) .
AE301-02 50 rxns20 l system
AE301-03 100 rxns20 l sytecm
EasyScriptFirst-Strand cDNA Synthesis SuperMix
Kit Components
Description
EasyScript First-Strand cDNA Synthesis SuperMix provides
all the necessary components for cDNA synthesis from total or
poly(A)+ RNA. The first-strand cDNA products generated by the
supermix are suitable for cloning, PCR or qPCR applications.
Reduced RNase H activity to reduce RNA template degradation
during the rst-strand cDNA synthesis.
Anchored oligo(dT)18 for higher yield and more full length cDNA.
cDNA up to 8 kb.
Appl ication
First strand cDNA synthesis.
AE301-02 AE301-03
EasyScriptRT/RI Enzyme Mix 50 l 250 l
2ES Reaction Mix 500 l 2500 l
Random Primer (0.1 g/l) 50 l 100 l
Anchored Oligo(dT)18 Primer (0.5 g/l) 50 l 100 l
RNase-free Water 500 l 1 ml
Storage
at -20oC for one year
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PROTOCOL
the First Strand cDNA synthesis1 Reaction Components
Component VolumeTotal RNA/mRNA Variable
Anchored Oligo(dT)18(0.5 g /l) 1 l
or Random Primer(N9) (0.1 g/l) 1 l
or GSP 2 pmol
2ES Reaction Mix 10 l
EasyScriptRT/RI Enzyme Mix 1 l
RNase-free Water to 20 l
2 Incubation
For anchored oligo(dT)18primer or GSP, incubate at 42oC for 30 min.
For random primer, incubate at 25oC for 10 min then at 42oC for 30
min.
3 Incubate at 85o
C for 5 min to inactive enzyme.RT-PCR
It is suggested to use 2-4 l reverse transcription products as PCR
templates.
The suggested reaction condition is the same as for EasyScript
Reverse Transcriptase ( P39 ).
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Kit Components
AT301-02 AT301-03
TransScriptRT/RI Enzyme Mix 50 l 250 l
2TS Reaction Mix 500 l 2500 l
Random Primer (0.1 g/l) 50 l 100 l
Anchored Oligo(dT)18 Primer (0.5 g/l) 50 l 100 l
RNase-free Water 500 l 1 ml
PROTOCOL
First Strand cDNA synthesis1 Reaction Components
Volume
Total RNA/mRNA Variable
Anchored Oligo(dT)18(0.5 g /l) 1 l
or Random Primer(N9) (0.1 g/l) 1 l
or GSP 2 pmol
2TS Reaction Mix 10 l
TransScript RT/RI Enzyme Mix 1 l
RNase-free Water to 20 l
2 Incubation
For anchored oligo(dT)18primer or GSP, incubate at 42oC for 30 min.
For random primer, incubate at 25
o
C for 10 min, then at 42
o
C for 30min.
3 Incubate at 85oC for 5 min to inactive enzyme.
RT-PCR
It is suggested to use 2-4 l reverse transcription products as PCR
templates.
The suggested reaction condition is the same as for EasyScript
Reverse Transcriptase( P39 ).
AT301-02 50 rxns20 l system
AT301-03 100 rxns20 l system
TransScript First-Strand cDNA Synthesis SuperMix
Description
TransScript First-Strand cDNA Synthesis SuperMix provides all the
necessary components for cDNA synthesis from total or poly(A)+ RNA.
The rst-strand cDNA products generated by this supermix are suitable
for cloning, PCR or qPCR applications.
Reduced RNase H activity to reduce RNA template degradation
during the rst-strand cDNA synthesis.
Anchored oligo(dT)18 for higher yield and more full length cDNA.
cDNA up to 12 kb.
Appl ication
First strand cDNA synthesis.
Storage
at -20oC for one year
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Description
Unique genomic DNA remover is combined with First-Strand cDNA
synthesis SuperMix to achieve simultaneous genomic DNA remove
and cDNA synthesis. After cDNA synthesis, gDNA remover and reverse
transcriptase are inactivated by heating at 85oC for 5 minutes.
Simultaneous genomic DNA removal and cDNA synthesis.
cDNA up to 12 kb.
Appl ication
First strand cDNA synthesis.
First Strand cDNA synthesis1 Reaction Components
2 Incubation
For anchored oligo(dT)18primer or GSP, incubate at 42oC for 30 min.
For random primer, incubate at 25oC for 10 min then at 42oC for 30
min.
3 Incubate at 85oC for 5 min to inactive enzyme.
RT-PCR
It is suggested to use 2-4 l reverse transcription products as PCR
templates.
The suggested reaction condition is the same as for EasyScript
Reverse Transcriptase( P39 ).
Kit Components
AT311-02 AT311-03
TransScript RT/RI Enzyme Mix 50 l 250 l
gDNA Remover 50 l 250 l
2TS Reaction Mix 500 l 2500 l
Random Primer (0.1 g/l) 50 l 100 l
Anchored Oligo(dT)18Primer (0.5 g/l) 50 l 100 l
RNase-free Water 500 l 1 ml
PROTOCOL
Volume
Total RNA/mRNA Variable
Anchored Oligo(dT)18 Primer (0.5 g/l) 1 l
or Random Primer (0.1 g/l) 1 l
or GSP 2 pmol
2TS Reaction Mix 10 l
TransScript RT/RI Enzyme Mix 1 l
gDNA Remover 1 l
RNase-free Water to 20 l
TransScript One-Step gDNA Removal and cDNA
Synthesis SuperMixAT311-02 50 rxns20 l system
AT311-03 100 rxns20 l sytecm
Storage
at -20oC for one year
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PROTOCOL
TransScriptAll-in-One First-Strand cDNA Synthesis
SuperMix for PCR
Storage
at -20oC for one year
DescriptionTransScript All-in-One First-Strand cDNA Synthesis SuperMix for
PCR provides all the necessary components for cDNA synthesis from
total or poly(A)+ RNA. The SuperMix is provided at 5 concentration
and used at 1concentration by adding template and H2O. cDNA
generated with the SuperMix are suitable for regular PCR. The kit is
not suitable for qPCR.
1-tube format for simple and fast setup and reduce pipetting
variability.
Fast 30 min cDNA synthesis protocol.
cDNA up to 12 kb.
Kit Components
First Strand cDNA synthesis
1. Reaction Components
2. Incubate at 42oC for 30 min.
3. Incubate at 85oC for 5 min to inactive enzyme.
RT-PCR
It is suggested to use 2-4 l reverse transcription products as PCR
templates.
The suggested reaction condition is the same as forEasyScript
Reverse Transcriptase( P39 ).
AT321-01
5TransScriptAll-in-One SuperMix for PCR 200 l
RNase-free Water 1 ml
Volume
Total RNA/mRNA 50 ng-5 g/5-500 ng
5TransScriptAll-in-One SuperMix for PCR 4 l
RNase-free Water to 20 l
AT321-01 50 rxns20 l system
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Order: Hot line:0086-010-51296890 0086-400-898-0321
TransScript All-in-One First-Strand cDNA Synthesis
SuperMix for qPCR
Storage
at -20oC for one year
DescriptionTransScript
All-in-One First-Strand cDNA Synthesis SuperMix for
qPCR provides all the necessary components for cDNA synthesis from
total or poly(A)+ RNA. The SuperMix is provided at 5 concentration
and used at 1concentration by adding template and H2O. cDNA
generated by the SuperMix are suitable for qPCR. The kit is not
suitable for regular PCR.
1-tube format for simple and fast setup and reduced pipetting
variability.
Fast 10 min cDNA synthesis protocol.
High compatibility with qPCR reagent.
Kit Components
First Strand cDNA synthesis1. Reaction Components
2. Incubate at 42oC for 10 min.
3. Incubate at 85oC for 5 sec to inactive enzyme.
qRT-PCRIt is suggested to use 2-4 l reverse transcription products as qPCR
templates.The suggested reaction condition is the same as forTransStartGreen
qPCR SuperMix ( P60 ).
AT331-01
5TransScriptAll-in-One SuperMix for qPCR 200 l
5TransScriptAll-in-One No-RT Control
SuperMix for qPCR20 l
RNase-free Water 1 ml
Volume
Total RNA/mRNA Variable
5TransScriptAll-in-One SuperMix for qPCR 4 l
RNase-free Water to 20 l
AT331-01 50 rxns 20 l system
PROTOCOL
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ter1 AH301-02 50 rxns20l system
AH301-03 100 rxns20 l sytecm
TransScript II First-Strand cDNA Synthesis SuperMix
Kit Components
DescriptionTransScriptII First-Strand cDNA Synthesis SuperMix provides all the
necessary components for cDNA synthesis from total or Poly(A)+ RNA.
It is an optimized cDNA synthesis kit withTransScriptII RT. First-Strand
cDNA can be synthesized at higher temperature resulting higher yield
and more full-length cDNA products.
Storage
at -20oC for one year
Reduced RNase H activity to reduce RNA template degradation
during the rst-strand cDNA synthesis.
Anchored oligo(dT)20 for higher yield and more full length cDNA.
cDNA up to 15 kb.
Appl icationFirst strand cDNA synthesis.
PROTOCOL
First Strand cDNA synthesis1 Reaction Components
2. Incubation
For anchored oligo(dT)20primer or GSP, incubate at 50
o
C for 30 min. For random primer, incubate at 25oC for 5-10 min, then at 50oC for
30 min.
For GC-rich or complex secondary structure RNA template, incubate
at 55oC, 30 min.
3. Incubate at 85oC for 5 min to inactive enzyme.
Component Volume
Total RNA/mRNA Variable
Anchored Oligo(dT)20(0.5 g /l) 1 l
or Random Primer(N9) (0.1 g/l) 1 l
or GSP 2 pmol
2TS II Reaction Mix 10 l
TransSciptIIRT/RI Enzyme Mix 1 l
RNase-free Water to 20 l
RT-PCRIt is suggested to use 2-4 l reverse transcription products as PCR
templates.
The suggested reaction condition is the same as for EasyScript
Reverse Transcriptase ( P39 ).
Components AH301-02 AH301-03
TransScript II RT/RI Enzyme Mix 50 l 250 l
2TS II Reaction Mix 500 l 2500 l
Random Primer(N9) (0.1 g/l) 50 l 100 l
Anchored Oligo(dT)20 Primer (0.5 g/l) 50 l 100 l
RNase-free Water 500 l 1 ml
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AH311-02 50 rxns20 l systemAH311-03 100 rxns20 l system
TransScript II One-Step gDNA Removal and cDNASynthesis SuperMix
Description
Unique genomic DNA remover is combined with First-Strand cDNA
synthesis SuperMix to achieve simultaneous genomic DNA remove
and cDNA synthesis. After cDNA synthesis, gDNA remover and
reverse transcriptase are inactivated by heating at 85oC for 5 minutes.
Simultaneous genomic DNA removal and cDNA synthesis.
cDNA up to 15 kb.
Storage
at -20oC for one year
Kit Components
First Strand cDNA synthesis1 Reaction Components
2. Incubation
For anchored oligo(dT)20primer or GSP, incubate at 50oC for 30 min.
For random primer, incubate at 25oC for 5-10 min, then at 50oC for
30 min.
3. Incubate at 85oC for 5 min to inactive enzyme.
RT-PCR
It is suggested to use 2-4 l reverse transcription products as PCR
templates.
The suggested reaction condition is the same as for EasyScript
Reverse Transcriptase( P39 ).
PROTOCOL
Volume
Total RNA/mRNA Variable
Anchored Oligo(dT)20 Primer (0.5 g/l) 1 l
or Random Primer (0.1 g/l) 1 l
or GSP 2 pmol
2TS II Reaction Mix 10 l
TransScript II RT/RI Enzyme Mix 1 l
gDNA Remover 1 l
RNase-free Water to 20 l
Components AH311-02 AH311-03
TransScript II RT/RI Enzyme Mix 50 l 250 l
gDNA Remover 50 l 250 l
2TS II Reaction Mi