ZymoTaq™ qPCR Premix

Hot-start polymerase for easy setup at room temperature reduces primer dimers.
Robust amplification of DNA for genotyping, SNP, and HRM analysis
Strong fluorescent signal for real-time and quantitative PCR assays

Product Size Catalog # Price Qty
ZymoTaq™ qPCR Premix 50 Rxns E2054
$90.00
ZymoTaq™ qPCR Premix 200 Rxns E2055
$335.00

About ZymoTaq™ qPCR Premix

The ZymoTaq™ qPCR PreMix product contains all the reagents needed to perform quantitative PCR and other molecular downstream analysis such as high-resolution melt (HRM) analysis of DNA methylation and other general real-time PCR assays.  It features a hot-start DNA polymerase and buffer system optimized for the amplification of bisulfite-treated DNA.  It also includes an intense dsDNA-specific fluorescent dye, SYTO 9®, for sensitive real-time DNA quantification. This technology can also be found in the Femto Quantification Kits (Cat. No. E2005, E2006, E2007) for low copy detection of DNA from human, bacteria, or fungi.


Storage Store at -20°C for up to 12 months. Minimize exposure to light. Avoid repeated freeze/thawing of reagents.
Unit Definition One unit (U) enzyme of ZymoTaq™ DNA Polymerase is defined as the amount of enzyme required for the incorporation of 10 nmol dNTPs into an acid-insoluble form in 30 minutes at 72°C.
Enzyme Concentration Reaction conditions at 1X (50 µl total volume) will contain 2 U of ZymoTaq™ DNA polymerase.

Real-Time Amplification Plot. This figure shows robust real-time amplification of 10 ng of bisulfite-converted human DNA using the ZymoTaq™ qPCR PreMix and primers designed to amplify a region of the MGMT promoter.

Reliable Standards for the Quantification of Human DNA: Human DNA Standards performed in duplicates 10-fold serial dilutions ranging from 20 ng to 20 fg.

Suggested Reaction Setup (50 µl):

Reagent

Volume

Final conc.

2X ZymoTaq™ qPCR PreMix

25 µl

1 X

Forward Primer (10 µM)

Variable

0.3 to 1 µM

Reverse Primer (10 µM)

Variable

0.3 to 1 µM

Template DNA

Variable

< 100 ng/20 µl

ddH20

to 50 µl

Total volume

50µl

 

Note:  The final concentration of MgCl2 in the reaction (above) is 2.75 mM.  If required, scale reaction reagent volumes accordingly to optimize the MgCl2, primer, and/or template concentrations.


Suggested Conditions for qPCR:

Set the real-time PCR instrument to excitation and emission wavelengths of 465 nm and 510 nm, respectively.

Note: A setting that is compatible with SYBRgreen® should be compatible with the SYTO 9® dye in this kit.

Initial denaturation

95 °C

10 min.

Denaturation

94 to 96 °C

30 sec.

Annealing

Variable

30-40 sec.

Extension

72 °C

30-60 sec. for ≤ 1kb*

 

30-40 Cycles

 

Final extension

72 °C

7 min.

Hold

4 °C

> 4 min.

*Note:  Add 15 to 30 seconds to the extension time for each additional kb over 1 kb.  Make adjustments to the temperature if necessary.

For specific notes and additional information, please see the product protocol PDF.
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