Protocol for bulk RNA sequencing of enriched human neutrophils from whole blood and estimation of sample purity.
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ABSTRACT:
SUBMITTER: Gonye ALK
PROVIDER: S-EPMC9946790 | biostudies-literature | 2023 Mar
REPOSITORIES: biostudies-literature
Publications
Protocol for bulk RNA sequencing of enriched human neutrophils from whole blood and estimation of sample purity.
Gonye Anna L K ALK LaSalle Thomas J TJ Freeman Samuel S SS Reyes Miguel M Hacohen Nir N Villani Alexandra-Chloé AC Sade-Feldman Moshe M
STAR protocols 20230223 1
Although neutrophils are the most abundant leukocyte in healthy individuals and impact outcomes of diseases ranging from sepsis to cancer, they remain understudied due to technical constraints of isolation, preservation, and sequencing. We present a modified Smart-Seq2 protocol for bulk RNA sequencing of neutrophils enriched from whole blood. We describe steps for neutrophil isolation, cDNA generation, library preparation, and sample purity estimation via a bioinformatic approach. Our approach p ...[more]
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