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PCR Related Products PCR Related Products
PCR Related ProductsPCR Related Products
Current ProductsCurrent Products Thermostable DNA polymerasesThermostable DNA polymerases
– PfuPfu– PfuTurboPfuTurbo– TaqPlus PrecisionTaqPlus Precision– TaqPlus LongTaqPlus Long– TaqTaq
TaqTaqExtenderExtender Perfect MatchPerfect Match Opti-PrimeOpti-Prime
Components of PCRComponents of PCR
Thermostable DNA polymeraseThermostable DNA polymerase Reaction BufferReaction Buffer PrimersPrimers Template DNATemplate DNA dNTPsdNTPs WaterWater
– Cycling ConditionsCycling Conditions
PCR QualitiesPCR Qualities
YieldYield FidelityFidelity SpecificitySpecificity LengthLength
Thermostable DNA Thermostable DNA polymerasespolymerases
PfuPfu PfuTurboPfuTurbo TaqPlus PrecisionTaqPlus Precision TaqPlus LongTaqPlus Long TaqTaq
error rate(x10 -6)
extensiontime
required(min/kbtarget)
RecommendedLength
vectortargets
PCR Target(kb)
genomictargets
PfuTurbo DNApolymerase
1.3 + 0.2 1
2
0-10
0-15
0-6
0-10
Pfu DNApolymerase
1.3 + 0.2 2 0-10 0-4
TaqPlusPrecisionsystem
3.9 + 1.3 1 0-15 0-10
TaqPlus Longsystem
7.6 + 1.2 0.5-1 0-35 0-18.5
Taq DNApolymerase
8.0 + 3.9 1 0-10 0-5
Functional Domains of Functional Domains of PfuPfu DNA DNA PolymerasePolymerase
5’-3’exo
3’-5’ exoDNA-dependent DNA polymerase
non-functional
proofreading DNA synthesisactivity
1 126 329 775
Fidelity AssayFidelity Assay
PCR PCR – not single round extensionnot single round extension
Mutations are detected in the Mutations are detected in the lacIlacI genegene
Color detection from Big Blue Color detection from Big Blue systemsystem
Results have been published in peer Results have been published in peer reviewed journalsreviewed journals
Characteristics of Characteristics of PfuPfu
High FidelityHigh Fidelity Native Hot Start activityNative Hot Start activity High ThermostabilityHigh Thermostability Slow polymerization rateSlow polymerization rate Available as native and cloned Available as native and cloned
enzymeenzyme
Accuracy of Polymerases Accuracy of Polymerases in PCRin PCR
Pfu
Dee
p V
ent
Ven
t
Ho
t Tu
b
Taq Tfl
Tb
r
Tm
a
0
100000
200000
300000
400000
500000
600000
700000
800000P
fu
Dee
p V
ent
Ven
t
Ho
t Tu
b
Taq Tfl
Tb
r
Tm
a
Extension TimeExtension TimePfuPfu requires 2 min/kb requires 2 min/kb
Taq
Pfu
Minutes of extension time/kb of PCR target
M 0.25 0.5 1 1.5 2 2.5 3 3.5 4
Activity vs Temperature Activity vs Temperature PfuPfu and and TaqTaq
percent activity
25 35 50 60 75 95
Incubation Temperature (degrees C)
PfuPfu: Hot Start Every Cycle: Hot Start Every Cycle
M T TS ATG TMg Pfu
Polymerase Polymerase ThermostabilityThermostability
DNA Polymerase Polymerization Opt Half Life 95C Pfu 70-80C 19 hours
Taq 70-80C 1 hour
Characteristics of Characteristics of PfuTurboPfuTurbo
Improved version of Improved version of PfuPfu Same high fidelity as Same high fidelity as PfuPfu Higher PCR product yield thanHigher PCR product yield than Pfu Pfu
alonealone Can use 1 min/kb extension timesCan use 1 min/kb extension times Works well with low amounts of Works well with low amounts of
DNA templateDNA template
PCR Enhancing Factor PCR Enhancing Factor (PEF)(PEF)
Isolated from Isolated from Pyrococcus Pyrococcus Lacks DNA polymerase and Lacks DNA polymerase and
exonuclease activitiesexonuclease activities Information on identity/mechanism Information on identity/mechanism
of action is proprietaryof action is proprietary
PfuTurboPfuTurbo vs Other vs Other PolymerasesPolymerases
M PT P V DV Pw K T M PT P V DV Pw K T M
0.9 kb
5.2 kb
PfuTurboPfuTurbo vs vs PfuPfu
M T P T P T P T P T P
0.9 kb
1.9 kb
8 kb
6.1 kb5.2 kb
Characteristics of Characteristics of TaqPlus PrecisionTaqPlus Precision
Blend of Blend of TaqTaq and and PfuPfu High YieldHigh Yield High FidelityHigh Fidelity LengthLength PCR Buffer PCR Buffer
– optimized for both yield and fidelityoptimized for both yield and fidelity
Accuracy of Polymerase Blends Accuracy of Polymerase Blends in PCRin PCR
020000400006000080000
100000120000140000160000180000
Taq
Plu
sP
reci
sio
n
Exp
and
Hig
hFi
de
lity
Exp
and
Lo
ng
Elo
ng
ase
LA
PC
R
Ad
van
tag
eK
len
Taq
Taq
Plu
s L
on
g
ExT
aq
Ad
van
tag
e T
th
Ge
ne
Am
p X
L
PCR with PCR with TaqPlus PrecisionTaqPlus Precision vs vs TaqTaq
Taq TaqPlus Precision M 2 5.2 5.4 10 15 M 2 5.2 5.4 10 15 kb
Characteristics of Characteristics of TaqPlus TaqPlus LongLong
Blend of Blend of TaqTaq and and PfuPfu Long targetsLong targets Fidelity similar to Fidelity similar to TaqTaq Two optimal buffers suppliedTwo optimal buffers supplied
– high salt buffer helps prevent high salt buffer helps prevent smearing in long target amplificationsmearing in long target amplification
– low salt buffer low salt buffer
PCR with PCR with TaqTaq vs vs TaqPlus TaqPlus LongLong
Qualities of Qualities of TaqTaq
Taq2000 Taq2000 is the recombinant is the recombinant TaqTaq Native Native TaqTaq is also available is also available TaqTaq is trusted by customers is trusted by customers TaqTaq has been well studied has been well studied
TaqTaqExtenderExtender
The The Pfu Pfu component of component of TaqPlus TaqPlus LongLong
Can be added to anyCan be added to any Taq Taq to make to make a polymerase blenda polymerase blend
Perfect Match PCR Perfect Match PCR EnhancerEnhancer Taq2000 native Taq
M - + - +
Opti-Prime PCR Optimization Opti-Prime PCR Optimization KitKit
12 buffers12 buffers– matrix of pH, Mg, KClmatrix of pH, Mg, KCl
AdjunctsAdjuncts– DMSO, formamide, BSA, glycerol, DMSO, formamide, BSA, glycerol,
ammonium sulfate, Perfect Matchammonium sulfate, Perfect Match Recommended for use with Recommended for use with TaqTaq
Related PCR productsRelated PCR products RoboCyclerRoboCycler
– gradientgradient Nucleotides and primers Nucleotides and primers StrataPrep PCR purification kitStrataPrep PCR purification kit Cloning VectorsCloning Vectors
– PCR-Script Kits PCR-Script Kits – Affinity protein expression kitAffinity protein expression kit
QuikChange site-directed mutagenesis kitQuikChange site-directed mutagenesis kit Cyclist DNA sequencing kitsCyclist DNA sequencing kits
