Digital RNA Sequencing

Digital RNA Sequencing

Digital RNA sequencing is an approach to RNA-Seq that removes sequence-dependent PCR amplification biases by barcoding the RNA molecules before amplification (Shiroguchi et al., 2012). RNA is reverse-transcribed to cDNA, and an excess of adapters, each with a unique barcode, is added to the preparation. This barcoded cDNA is amplified and sequenced. Deep sequencing reads are compared, and the barcodes are used to determine the actual distribution of RNA abundance.

Advantages:

• Low amplification bias during PCR
• Provides information about abundance of RNA
• Detection of low-copy_number RNA
• Single-copy resolution

Disadvantages:

• Some amplification bias still persists
• Barcodes may miss targets during ligation

Reagents:

Illumina Library prep and Array Kit Selector



Reviews:

This method has been widely integrated into various sequencing techniques due to its high versatility.

References:

This method has been widely integrated into various sequencing techniques due to its high versatility.

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History: Digital RNA Sequencing

Revision by sbrumpton on 2017-06-21 07:50:22 - Show/Hide