tRNAs carry the largest number and the most chemically diverse post-transcriptional modifications of any RNA species, and these marks seriously hamper tRNA cDNA synthesis. Methylation is the most common culprit: it blocks transcriptional elongation, resulting in poor cDNA conversion and unreliable qPCR readouts.
Arraystar solved this problem with an excellent RNA demethylase. The rtStar tRNA-optimized First-Strand cDNA Synthesis Kit (Cat# AS-FS-004) removes methylations from RNA before reverse transcription, delivering a new level of accuracy for tRNA detection.
Fig. 1. tRNA demethylation dramatically improves PCR assay performance. Upper: tRNA levels in three pancreatic carcinomas and three matched para-carcinoma RNA samples with or without demethylation (error bars, SEM, n=3; De, demethylated; Un, untreated). Bottom: principal component analysis of tRNA profiles. Demethylated cancer and para-carcinoma samples separate clearly, whereas untreated samples are indistinguishable.
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rtStar tRNA-optimized First-Strand cDNA Synthesis Kit
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