tRNAs undergo by far the greatest number of and the most chemically diverse post-transcriptional modifications, which badly affect tRNA cDNA synthesis efficiency. Methylation, one of the most prevalent modifications, is the main obstacle that hinders transcriptional elongation and thus leads to non-effective cDNA conversion and qPCR. By using an excellent RNA demethylase, Arraystar developed rtStar™ tRNA-optimized First-Strand cDNA Synthesis Kit (Cat# AS-FS-004), a highly efficient system that removes methylations on the RNA to obtain a new level of accuracy on the tRNA detection.
Figure 1. tRNA demethylation treatment dramatically increased PCR assay performance. Upper: tRNA levels in 3 pancreatic carcinomas and 3 matched para-carcinoma RNA samples with or without the RNA demethylation treatment. Error bars indicate SEM (n=3). De: demethylation, Un: untreated with demethyation. Bottom: Principle component analysis (PCA) analysis of tRNA profiles for the pancreatic carcinoma and para-carcinoma tissues. The Cancer (Demethylation) and Para-carcinoma (Demethylation) samples are well separated, whereas the Cancer and Para-carcinoma samples without the demethylation treatment are indistinguishable.
rtStar™ tRNA-optimized First-Strand cDNA Synthesis Kit
nrStar™ tRNA PCR Arrays