RT_ Easy First Strand Kit (12)

From QIAGEN, the RT² Easy First Strand Kit is designed for cDNA synthesis and genomic DNA elimination in RNA samples intended for use with RT² Profiler PCR Arrays. It provides a rapid and convenient procedure for efficient first strand cDNA synthesis and elimination of contaminating genomic DNA, with a format that enables easy parallel processing of multiple RNA samples for reverse transcription.

Random hexamers and oligo-dT primers prime reverse transcription in an unbiased manner, while a reverse transcriptase synthesizes cDNA with optimal yield and length. A built-in external RNA control helps monitor reverse transcription efficiency and test for the presence of enzyme inhibitors. The proprietary Buffer GE2 is mixed with the sample and incubated on a thermal cycler; after addition of enzyme and additional buffer, the sample is reverse transcribed on any thermal cycler, and the resulting cDNA is immediately ready for downstream applications such as real-time PCR expression analysis with RT² SYBR Green Mastermixes, RT² qPCR Primer Assays, or other commercially available primer assays.

The kit handles as little as 25 ng or as much as 5 µg total RNA per reaction, with the optimal amount depending on the relative abundance of the transcripts of interest. It is optimized for gene expression analysis using RT² Profiler PCR Arrays and RT² qPCR Primer Assays, and the chemistries are not compatible with AMBION's Turbo DNA-Free Kits. We also recommend using the RNeasy Mini Kit with RNase-free DNase, or kits such as RNeasy Plus Universal Tissue which include a DNA removal step.