The Deoxy+ OneStep RT-PCR Kit is a ready-to-use master mix which eliminates the need for optimization of reaction and cycling conditions for one-step RT-PCR. The reaction can be prepared by simply adding template RNA and primers to the master mix. The use of Yeastern’s Hotstart DNA polymerase and Deoxy+ HiSpec RT enables reliable real-time RT-PCR quantification on any real-time PCR machines. Since it is a one-tube reaction, the procedure makes high-throughput analysis possible.
After reverse transcription, reactions are heated to 95°C for 10 minutes to inactivate the reverse transcriptase and simultaneously activate HotStart Taq DNA polymerase. This hot start to the PCR eliminates any nonspecific amplification products such as primer-dimers and reduces background smear, ensuring highly sensitive and reproducible RT-PCR.
- Fast and easy one-tube setup
- One-step RT-PCR of any RNA template without optimization
- Unique enzyme mix for high specificity and sensitivity
- Optimized reverse-transcription and amplification buffer
- Single-cell RT-PCR
- Gene-expression analysis
The performance of Deoxy+ OneStep RT-PCR Kit is tested in an RT reaction using human embryonic kidney cell lysate with primer d(T)18. The sensitivity of the kit is verified by the detection of B2M transcript after 35 cycles. The length of cDNA achieved is verified by detection of a 248 bp by DNA agarose gel and DNA sequencing.