使用条码测序分析在酵母中进行批量健身测量。

Bulk-Fitness Measurements Using Barcode Sequencing Analysis in Yeast.

机构信息

Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada.

Department of Biology, University of Toronto at Mississauga, Mississauga, ON, Canada.

出版信息

Methods Mol Biol. 2022;2477:399-415. doi: 10.1007/978-1-0716-2257-5_22.

DOI:10.1007/978-1-0716-2257-5_22

PMID:35524129

Abstract

The use of DNA barcodes for determining changes in genotype frequencies has been instrumental to increase the scale at which we can phenotype strain libraries by using next-generation sequencing technologies. Here, we describe the determination of strain fitness for thousands of yeast strains simultaneously in a single assay using recent innovations that increase the precision of these measurements, such as the inclusion of unique-molecular identifiers (UMIs) and purification by solid-phase reverse immobilization (SPRI) beads.

摘要

DNA 条码在确定基因型频率变化方面的应用对于提高我们利用下一代测序技术对菌株文库进行表型分析的规模起到了关键作用。在这里，我们描述了使用最近的创新技术，如包含独特分子标识符 (UMI) 和固相反向固定 (SPRI) 珠纯化，在单个测定中同时确定数千个酵母菌株的菌株适应性的方法。这些测量方法的精度得到了提高。