from Bits of DNA by Lior Pachter
Today my student Harold Pimentel released the beta version of his new RNA-Seq analysis method and software program called sleuth. A sleuth for RNA-Seq begins with the quantification of samples with kallisto, and together a sleuth of kallistos can be used to analyze RNA-Seq data rigorously and rapidly.
Why do we need another RNA-Seq program?
A major challenge in transcriptome analysis is to determine the transcripts that have changed in their abundance across conditions. This challenge is not entirely trivial because the stochasticity in transcription both within and between cells (biological variation), and the randomness in the experiment (RNA-Seq) that is used to determine transcript abundances (technical variation), make it difficult to determine what constitutes “significant” change.
Technical variation can be assessed by performing technical replicates of experiments. In the case of RNA-Seq, this can be done by repeatedly sequencing from one cDNA library. Such replicates are fundamentally distinct from biological replicates designed to assess biological variation. Biological replicates are performed by sequencing different cDNA libraries that have been constructed from repeated biological experiments performed under the same (or in practice near-same) conditions. Because biological replicates require sequencing different cDNA libraries, a key point is that biological replicates include technical variation.