High-throughput Tetrad Analysis

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2013 May 14

PMID 23666411

Citations 13

Authors

Catherine L Ludlow

Adrian C Scott

Gareth A Cromie

Eric W Jeffery

Amy Sirr

Patrick May

Jake Lin

Teresa L Gilbert

Michelle Hays

Aimee M Dudley

Affiliations

Soon will be listed here.

Abstract

Tetrad analysis has been a gold-standard genetic technique for several decades. Unfortunately, the need to manually isolate, disrupt and space tetrads has relegated its application to small-scale studies and limited its integration with high-throughput DNA sequencing technologies. We have developed a rapid, high-throughput method, called barcode-enabled sequencing of tetrads (BEST), that uses (i) a meiosis-specific GFP fusion protein to isolate tetrads by FACS and (ii) molecular barcodes that are read during genotyping to identify spores derived from the same tetrad. Maintaining tetrad information allows accurate inference of missing genetic markers and full genotypes of missing (and presumably nonviable) individuals. An individual researcher was able to isolate over 3,000 yeast tetrads in 3 h, an output equivalent to that of almost 1 month of manual dissection. BEST is transferable to other microorganisms for which meiotic mapping is significantly more laborious.

Citing Articles

Applications of Programmable Endonucleases in Sequence- and Ligation-Independent Seamless DNA Assembly.

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A method for quantifying sporulation efficiency and isolating meiotic progeny in non-GMO strains of Saccharomyces cerevisiae.

Sirr A, Timour M, Cromie G, Tang M, Dudley A Yeast. 2022; 39(6-7):354-362.

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Computational Inference Software for Tetrad Assembly from Randomly Arrayed Yeast Colonies.

Sakhanenko N, Cromie G, Dudley A, Galas D G3 (Bethesda). 2019; 9(7):2071-2088.

PMID: 31109921 PMC: 6643883. DOI: 10.1534/g3.119.400166.

Rapid Phenotypic and Genotypic Diversification After Exposure to the Oral Host Niche in .

Forche A, Cromie G, Gerstein A, Solis N, Pisithkul T, Srifa W Genetics. 2018; 209(3):725-741.

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