FAST qPCR 2x Master Mix (probe, UDG)  includes all kind of real-time qPCR components such as hot start Taq DNA polymerase, dNTP and buffer. The only need to perform the reactions template DNA, Taqman probes and primers provided by users.

   Especially, this product is suitable to perform the Taqman real-time qPCR reaction, since it has Taq DNA polymerase having the 5’ to 3’ exonuclease activity. In addition, Taq polymerase is a mixture of PCR enzyme and monoclonal antibody to Taq DNA polymerase. The antibody inhibits the polymerase activity and prevent non-specific amplification derived from mispriming of primer dimer before PCR cycling.

 

Characteristics

 

  Enhanced specificity by use of HOT start with anti-Taq antibody 

 

Qualifying Test

 

  Performance test, exo/endo nuclease contamination test, stability test. 

 

Storage Condition

 

  For long term storage, we recommend to store at -20℃ (stable up to 6 months), and store at 4℃ after thawing the solution instead of -20℃. 

 

Applications

 

- Real-Time quantification of DNA and cDNA targets

- Gene expression profiling.

- Microbial & viral pathogen detection.

- Diverse applications using qPCR

- Multiplex PCR

 

Standard Experiment Protocol

1. Thaw the FAST qPCR 2x Master Mix.

2. Prepare the 20㎕ of qPCR reaction mix as follow.

 

FAST qPCR 2x Master Mix                          10 ㎕ 

Primer up-stream (10 pmol / ㎕)            0.5 - 1 ㎕

Primer down-stream (10 pmol / ㎕)       0.5 - 1 ㎕

Probe (10 pmol / ㎕)                           0.2 - 0.5 ㎕

Template DNA  (1-100 ng)                      variable

 

Adjust final vol. to 20 ㎕ with PCR grade distilled water

 

3. After mix and put the tubes or plates.

4. Seal the tubes or plates with cap or optical adhesive film.

5. Run the Real-Time qPCR machine using the program.

Initial denaturation	95 ℃	1-3 min
Denaturation  Annealing & Extension	95 ℃  60 ℃	10-20 sec  20-60 sec	40-50 cycles

    a. The number of PCR cycle is dependent on the copy number of DNA target in the samples.

    b. Optimal annealing temperature is decided by the melting temperature of primer pairs. Usually, use the  2-4℃ below than the melting points of primer pairs.

    c. The precise adjustment of experimental procedure should be performed according to the samples and purpose.

6. Analysis the results.