I am going to buy GoTaq® 2-Step RT-qPCR System(Promega). This kit uses oligo(dT)15 primer for first-strand cDNA synthesis. I have to design primers for qPCR for 4 mature microRNAs(miR-21, miR-155, miR-205, let-7b).
And is there anybody here used this kit for microRNA qPCR? What is your opinion about the performance?
MicroRNA is very small RNA and to perform PCR for it you should elongate it using universal primer then use a single specific primer and a primer complementary to the universal sequence. oligo dT is a small TTTTTT sequence which anneal to the terminal AAAAA sequence usually in mRNA. So not suitable for your work.
My advise use a complete kit for microRNA from a reputed company which has a reverse transcription kit for microRNA e.g. Qiagen miscript RT with the universal primer included. and microRNA assay for your target genes with the sybergreen mastermix
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