title: High-throughput tetrad analysis.
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.
authors.affiliation: Institute for Systems Biology, Seattle, Washington, USA.
authors.forename: Catherine L
authors.initials: CL
authors.lastname: Ludlow
authors.forename: Adrian C
authors.initials: AC
authors.lastname: Scott
authors.forename: Gareth A
authors.initials: GA
authors.lastname: Cromie
authors.forename: Eric W
authors.initials: EW
authors.lastname: Jeffery
authors.forename: Amy
authors.initials: A
authors.lastname: Sirr
authors.forename: Patrick
authors.initials: P
authors.lastname: May
authors.forename: Jake
authors.initials: J
authors.lastname: Lin
authors.forename: Teresa L
authors.initials: TL
authors.lastname: Gilbert
authors.forename: Michelle
authors.initials: M
authors.lastname: Hays
authors.forename: Aimée M
authors.initials: AM
authors.lastname: Dudley
mesh_terms: D000465:Algorithms; D002874:Chromosome Mapping / Q000379:methods*; D004271:DNA, Fungal / Q000235:genetics*; D005819:Genetic Markers / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D008540:Meiosis / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*
chemical_list: D004271:DNA, Fungal; D005819:Genetic Markers