RNA Sequencing (RNAseq)

A misnomer of sorts, RNA sequencing (or “RNAseq”) does not directly sequence RNA. Rather, RNA is first converted to complementary DNA (or “cDNA”) and this material is then sequenced. RNAseq is most often used to gain insight into gene expression. The reads from the cDNA are used to determine the relative abundance of various RNA constructs in a given sample.

For mRNA sequencing, poly A enrichment using an oligo(dT) primer is used to target just the mRNAs. For Total RNA sequencing, random priming is used for first strand cDNA synthesis and reads from all of the various RNAs can be produced (e.g. lncRNA, tRNA, mRNA, rRNA).

The random priming process performed in the standard Total RNA sequencing workflow tends to produce a preponderance of ribosomal RNA (rRNA) in the resulting data. If this is a problem for your research, rRNA depletion/removal can be performed for an extra cost per sample.

Data is delivered as FASTQ.gz files through Globus Connect or a direct transfer into your Amazon Web Services (AWS) S3 account, if available.

RNA Sequencing (RNAseq)

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