VNIRTM Green 2X One-step RT-qPCR Master Mix (ROX)

All Products Molecular Biology Products VNIR 115

Description

VNIRTM Green 2X One-step RT-qPCR Master Mix is based on quantitative reverse transcription PCR (RT-qPCR) that uses RNA as the starting material. It offers a convenient master mix to convert RNA to DNA and quantify in a one-step reaction. The single-tube 2X formulation contains MMLV reverse transcriptase, dNTPs, VNIR Green (equivalent to Sybr® Green), Taq DNA polymerase, reaction buffer, MgCl2, KCl, and PCR enhancer/stabilizer facilitating easy reaction set up. The user needs only to add the template, the primer set and water to the master mix to set up the RT-PCR reaction. This master mix is useful for efficient amplification of GC rich, low copy number genes from various template types and can be used up to 80 cycles. This master mix contains 2.5mM MgCl2 (1X final concentration).

Cat. No.: VNIR115

Quality Control

Assay
Results
Amplification efficiency >105 fold
qPCR reproducibility Passed
ds Endodeoxyribonuclease activity Not observed
ss Exodeoxyribonuclease activity Not observed
Self-priming activity Not observed

Recommended reaction setup conditions

Component
Volume
Final Concentration
VNIR Green 2X One-step RT-qPCR Master Mix 10µl 1X
Forward Primer – 10µM 0.5µl 500nM
Reverse Primer – 10µM 0.5µl 500nM
RNA Template Variable 50-400ng
PCR grade water Variable
Total Volume per reaction 20 µl

*vortex briefly and spin down after addition of all components to ensure complete mixing

Amplification Conditions

Step
Temperature
Time
Cycles
RT 42°C 5-10 min 1
Enzyme Activation 95°C 3 min 35-45
Denaturation 95°C 30 sec
Annealing, extension and read fluorescence Primers Tm 30 sec
Storage conditions
-20°C (protect from light)
Best before
24 months upon receiving
References
Please let us know if you publish your research using VNIRTM Green 2X One-step RT-qPCR Master Mix. We can cite your reference here.
Note
This formulation doesn’t contain ROX.