DETERMINING THE ACCURACY OF TRANSCRIPT QUANTIFICATION BY RNA-SEQ

 

Jerod Parsons, P.Scott Pine, Marc Salit, Sarah Munro, Jennifer McDaniel

 

The burgeoning field of RNA-seq has opened the door to making high throughput measurements of complete transcriptomes, but there are open questions regarding the best way to interpret the raw sequencing data.  To date, there has been only low-throughput sampling of the accuracy of individual measurements by qPCR.  By comparing several pure tissue samples to complex mixtures containing fixed ratios of these pure samples, the accuracy of thousands of measurements of RNA transcripts can be determined at once.

 

Several out of over a dozen commonly used practical methods and implementations for quantifying transcripts and identifying differential expression are evaluated for accuracy, sensitivity, and specificity by using these mixtures.