FYT108-100PFYT108-400P

Cat. No.

(2X, For SYBR® Green, ROX)

Fast Real-Time PCR Premix

Ver. L0524

(2X, For SYBR® Green, ROX)

EZtime™ Fast Real-Time PCR Premix

Description

Content

Applications

Storage

Procedure

EZtime™ Fast Real-Time PCR Premix for SYBR® Green is a ready-to-use, 2X concentrated premix reagent, containing all components except primers and template. It is formulated with a novel hot-start Taq DNA polymerase, which is capable of catalyzing DNA amplification in a fast PCR mode. This special blend greatly shortens the running time of real-time quantitative PCR by around 1 hour when compared to traditional qPCR. In addition, it precisely meets current researchers’ needs for performing gene detection (qPCR) and quantification of gene expression (2-step qRT-PCR) in a high speed and/or high-throughput manner in addition to those basic requirements of high sensitivity, wide dynamic range, and good reproducibility.

NoteFor research only. Not for use in diagnostic or therapeutic procedures.

Figure 1

Quantitative real-time PCR for DNA template below 300 bp. Gene expression analysisLow copy gene detectionGenotyping in a single or high-throughput mannerValidation of microarray data

1. Thaw all reagents including EZtime™ Fast Real-Time PCR Premix, primers and DNA template.2. Prepare a PCR master mixture according to Table 1.3. Mix carefully by pipetting up and down.

-20°CProtected from lightAvoid repeated freezing and thawing

A. Preparing the reaction mixture on ice

1. Program your instrument according to Figure 1.2. Place the PCR tubes or PCR plates in the thermo cycler and start the cycling program.

3. Perform a melting curve analysis of the PCR product.4. Perform data analysis.

B. Performing Real-time PCR

EZtime™ Fast Real-TimePCR 2x Premix for SYBR® Green 10

0.6-1.2 0.3-0.6 µM10 µM Forward Primer

0.6-1.2

variable

0.3-0.6 µM10 µM Reverse Primer

2Template DNA

ddH2O

Component Volume (µl) Final conc.

1x

95 oC

20 sec 3 sec

95oC

4oC≥ 20 sec

58-65 oC

∞40-45 cycles1 cycle

Table 1

FYT108-100P EZtime™ Fast Real-Time PCR 2X Premix with SYBR®

Green 1 ml*1 vialFYT108-400P EZtime™ Fast Real-Time PCR 2X Premix with SYBR® Green 1 ml*4 vials

20 µlTotal volume

FYT110-100PFYT110-400P

Cat. No.

(2X, For TaqMan® Probe, ROX)

Fast Real-Time PCR Premix

Ver. L0524

(2X, For TaqMan® Probe, ROX)

EZtime™ Fast Real-Time PCR Premix

Description

Content

Applications

Storage

Procedure

EZtime™ Fast Real-Time PCR Premix for TaqMan® Probe is a ready-to-use, 2X concentrated premix reagent, containing all components except primers, probe, and template. It is formulated with a novel hot-start Taq DNA polymerase, which is capable of catalyzing DNA amplification in a fast PCR mode. This special blend greatly shortens the running time of real-time quantitative PCR by around 1 hour when compared to traditional qPCR. In addition, it precisely meets current researchers’ needs for performing gene detection (qPCR) and quantification of gene expression (2-step qRT-PCR) in a high speed and/or high-throughput manner in addition to those basic requirements of high sensitivity, wide dynamic range, and good reproducibil-ity.

FYT110-100P EZtime™ Fast Real-Time PCR 2X Premix with TaqMan® Probe 1 ml *1 vialFYT110-400P EZtime™ Fast Real-Time PCR 2X Premix with TaqMan® Probe 1 ml*4 vials

NoteFor research only. Not for use in diagnostic or therapeutic procedures.

Table 1

Figure 1

Quantitative real-time PCR for DNA template below 300 bp. Gene expression analysisLow copy gene detectionGenotyping in a single or high-throughput mannerValidation of microarray data

1. Thaw all reagents including EZtime™ Fast Real-Time PCR Premix, primers, probe and DNA template.2. Prepare a PCR master mixture according to Table 1.3. Mix carefully by pipetting up and down.

-20°CProtected from lightAvoid repeated freezing and thawing

A. Preparing the reaction mixture on ice

1. Program your instrument according to Figure 1.2. Place the PCR tubes or PCR plates in the thermo cycler and start the cycling program.

3. Perform data analysis.

B. Performing Real-time PCR

EZtime™ Fast Real-Time PCR2x Premix for TaqMan® Probe 10

0.6-1.2 0.3-0.6 µM10 µM Forward Primer

0.6-1.2 0.3-0.6 µM

0.4-0.8 0.2-0.4 µM

10 µM Reverse Primer

10 µM Probe

Component Volume (µl) Final conc.

1x

95 oC

20 sec 3 sec

95oC

4oC≥ 20 sec

58-65 oC

∞40-45 cycles1 cycle

variable

2Template DNA

ddH2O

20 µlTotal volume