Product Description
The 2x Foci-Fast PCR master mix contains the engineered next generation Taq DNA polymerase that is at least three times faster than regular Taq.
The master mix is a pre-mixed 2x solution of fast Taq enzyme, PCR reaction buffer, dNTPs, MgCl2, enzyme stabilizer, PCR enhancers and SYBR® Green fluorescein. It contains all the necessary components for qPCR amplification except template DNA and primers. This PCR master mix has been optimized for high-efficiency qPCR amplification, and is ideal for quantifying cDNA and genomic DNA.
Features:
Super fast, the Foci-Fast next generation DNA polymerase in the master mix significantly reduced the time required for qPCR reaction.
Enhanced Primer-template binding.
Ultra sensitive, can detect trace amount of genomic DNA or cDNA.
Affordable, significantly cheaper than the similar products from competitors.
Storage:
For long term storage, keep at -80°C or -20°C. Shield against light. For short term storage, thaw the master mix and store at 4°C. The master mix is stable at 4°C for at least one month. Avoid repeated freeze and thaws.
Fig. 1. The cDNA synthesized using Gene-Foci® High Efficiency Reverse transcription kit was diluted 1:200, 1:500, 1:2000 and 1:5000, then GAPDH expression level was examined with Foci-Fast SYBR® Green qCR Master Mix.
qPCR Reaction Setup
To setup a 25µl qPCR reaction, add the following components into a DNase-Free qPCR plate or tube:
|
Components |
Volume |
Final Concentration |
|
2X Fast PCR Master Mix |
12.5 µl |
1X |
|
Forward Primer (1 μM) |
1~7.5 µl |
50~300 nM[1] |
|
Reverse Primer (1 μM) |
1~7.5 µl |
50~300 nM[1] |
|
DNA Template |
1~5 µl |
0.1~10 ng/μl |
|
ddH2O |
to 25 µl |
[1] A primer titration from 50 nM to 300 nM final concentration should be performed to determine the most amplification-efficient concentration.
qPCR Thermal Cycling Program
Set up the program qPCR on a qPCR machine as follows:
Option I: Two-step PCR
|
Cycles |
Temperature |
Time |
|
| (1) Pre-denature |
1 |
95°C |
3 minutes |
| (2) Thermal cycling |
40 |
95°C |
10 seconds |
|
55~60°C(a) |
15~20 seconds |
Option II: Three-step PCR
|
Cycles |
Temperature |
Time |
|
| (1) Pre-denature |
1 |
95°C |
3 minutes |
| (2) Thermal cycling |
40 |
95°C |
10 seconds |
|
55~60°C(a) |
10 seconds |
||
|
72°C |
10~15 seconds |
(a) Temperature required for annealing/extension step may need adjustment for different DNA templates and primers.
