The Sensiscript RT Kit includes dNTPs and an optimized reaction buffer that, together with the high affinity of Sensiscript Reverse Transcriptase for RNA, enables read-through of templates with high GC content or complex secondary structures. Due to this pre-optimization, tedious pipetting and pre-incubation steps are eliminated, and no additional RNase H digestion step is required.
Total RNA was isolated from 10 to 5000 HeLa cells using RNeasy technology. One-tenth of each eluate (corresponding to RNA from 1 to 500 cells) was used for reverse transcription with Sensiscript Reverse Transcriptase. One-half of each RT reaction was then used in a 40-cycle PCR with QIAGEN Taq DNA Polymerase and primers specific for genes encoding β-actin, NF-κB, inositol-1,4,5-triphosphate receptor, or dystrophin. Sizes of the amplicons are as indicated.