Member since: 2019
Organization: University of Manitoba
Easy to use for cDNA generation and great qPCR results.
Application Area: Reverse transcription of RNA to cDNA
"I used purified total RNA from whole mouse brain as my template. Following the protocol as outlined from NEB, I used 1ug of RNA as template for the reverse transcription. The protocol was simple and straight forward, requiring <2 minutes hands-on time and only 15 minutes on a thermocycler. I purified my cDNA using a PCR cleanup kit and analyzed my cDNA on a NanoDrop 2000. I had great 260/280 and 260/230 ratios, with a total recovery of approximately 70% of input RNA. I diluted this cDNA to 10ng/uL, then a 2-fold dilution curve and tested amplification on by qPCR. The ct values were as expected, and the amplification followed the dilutions. Note that I did an on-column DNase digestion during RNA purification and use exon-exon spanning primers in order to negate gDNA amplification. I recommend LunaScript RT SuperMix for generation of single stranded cDNA. Simple to use with good output."
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