PACE OneStep RT-PCR Master Mix combines reverse transcription (RT) and PCR in a single reaction. This advanced master mix simplifies workflows by eliminating the need for separate reactions for reverse transcription of RNA to cDNA, and PCR amplification from the newly generated cDNA. PACE OneStep RT-PCR Master Mix is highly efficient and sensitive, enabling detection of low-abundance RNA targets, particularly useful for applications such as gene expression analysis and viral RNA detection. PACE OneStep RT-PCR Master Mix demonstrates robust performance across a wide range of RNA templates and can be employed for both routine and challenging samples.
PACE OneStep RT-PCR Master Mix is designed for highly specific and sensitive one-step reverse transcriptase (RT) fluorescently-reporting endpoint PCR using RNA template.
PACE OneStep RT-PCR Master Mix includes optimised components which allow reverse transcription and subsequent PCR amplification to take place in the same reaction well.
The PACE OneStep RT-PCR reaction is comprised of three parts:
When combined with template RNA, these components create a PACE OneStep RT-PCR.
A one-step RT-PCR (Reverse Transcription Polymerase Chain Reaction) allows for the amplification of a target DNA sequence directly from an RNA sample. It combines reverse transcription of RNA into complementary DNA (cDNA) and subsequent amplification of the target DNA sequence in a single reaction tube. Reverse transcription and cDNA amplification can also be carried out in two separate reactions (two-step method), but the one-step approach significantly improves and streamlines the workflow and reduces the risk of contamination.
One-step RT-PCR combines reverse transcription of RNA into complementary DNA (cDNA) and subsequent amplification of the target DNA sequence in a single reaction tube. Reverse transcription and cDNA amplification can also be carried out in two separate reactions (two-step method), but the one-step approach significantly improves and streamlines the workflow and reduces the risk of contamination.
PACE OneStep RT-PCR Master Mix can be used with any standard qPCR instrument, peltier or water bath-based thermal cycler. The fluorescent data should be captured at the PCR endpoint at 40˚C or below.
PACE OneStep RT-PCR Master Mix can be used with any reaction plate or well volume. It is important that PACE OneStep RT-PCR Master Mix is used at a final 1x concentration.
RNA is highly susceptible to ubiquitous RNases. Care should be taken when handling the samples. An empirical optimisation of RNA concentration should be carried out by testing a sample dilution range.
If RNA samples contain EDTA, the concentration at the final reaction concentration should be no higher than 0.1 mM.
This should be considered when using PACE OneStep RT-PCR Master Mix.
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