Year: 2018
Type: article
Abstract: ScarTrace is a protocol that allows the simultaneous ampli cation of the transcriptome and Cas9induced genetic scars of thousands of zebra sh single cells.Initially, Tg\(H2Af/va-GFP)\(kca66) zebra sh one-cell embryos are injected with Cas9 RNA or protein and a gRNA targeting a GFP transgene to allow scaring.At adulthood, various zebra sh organs are dissected, dissociated and single cells are sorted.The following protocol describes the steps to generate transcriptome and scar sequencing libraries from single cells from the WKM, brain, eyes and caudal n.Brie y, ScarTrace is an adaptation of the SORT-Seq protocol1 where a nested PCR step was integrated to amplify scars after mRNA conversion to cDNA.
Source: Protocol Exchange
Authors: Chloé Baron, Josi Peterson-Maduro, Maria Florescu, Chloé S Baron, Anna Alemany, Alexander van Oudenaarden
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
OpenAlex ID: W2808118938