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URL: https://dx.doi.org/DOI:10.17504/protocols.io.y9gfz3w

Authors: Amy Lyden, Emily Crawford, Jenai Quan, Saharai Caldera, David Dynerman

Group: Chan Zuckerberg Biohub

Summary: This protocol is for performing Depletion of Abundant Sequences by Hybridization (DASH) after preparing sequencing libraries and pooling together.DASH is most useful for RNA-seq of human metagenomic samples, where abundant species such as human mitochondrial ribosomal RNAs (rRNAs) occupy a majority of the sequencing space available, leaving a minor fraction for regions of interest. DASH treatment is performed after ligation of adapters and unique barcoding of the RNA-seq library. It employs CRISPR-Cas9 complexed to a set of guide RNAs (gRNAs) targeted to the abundant regions to be depleted in a given library. These abundant regions in the library are then cleaved, leaving only the fragments with intact adapters on both ends to be further amplified and sequenced.

Affiliations: CZ Biohub, CZ Biohub, CZ Biohub, UCSF, CZ Biohub

Version: 1

Publication Date: 2019

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