One-step RT-PCR 2x Premix is a mixture of thermostable DNA Polymerase, thermostable M-MLV reverse transcriptase, dNTPs, reaction buffer, and stabilizer. Users can perform RT-PCR by simply adding total RNA or poly(A) RNA as substrates and specific primer pair to the premix. One-step RT-PCR 2x Premix also contains glycerol and bromophenol blue as a tracking dye, enabling a direct analysis of PCR products on an analytic gel electrophoresis. Stabilizer in premix enables prolonging a shelf life at -20℃. You do not need to dissolve the mixture prior to RT-PCR reaction as does competitor’ freeze-dried one. The composition is optimized to reduce non-specific amplification and to minimize primer-dimer formation.

 

 

Application(s) 

One-step RT-PCR for gel analysis.

 

 

Reagents Supplied 

One-step RT-PCR 2x Premix, 10㎕ in 12 x 8 strip tubes.

 

 

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).

 

 

Standard Experiment Protocol

1. Thaw the One-step RT-PCR 2x Premix

 

2. Prepare the 20 ㎕ of RT-PCR reaction mix as follow.

 

One-step RT-PCR 2x Master Mix                    10 ㎕

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

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

Template RNA (1 – 100 ng)                        variable

                                                                                   

Add distilled water to make 20 ㎕ final volume.

 

3. After mix and put the tubes on the PCR machine.