Transfer of YAC clones to new hosts by karyogamy-deficient mating.

F. Spencer, G. Simchen

Research output: Contribution to journalArticlepeer-review

Abstract

This unit provides a protocol for moving yeast artificial chromosome (YAC) clones to new yeast hosts using basic microbial techniques and pulsed-field gel analysis. In contrast to other methods that can be used to transfer YAC clones, this requires neither optimization to achieve high-efficiency DNA-mediated transformation of chromosome-sized DNA nor specialized equipment for tetrad dissection and analysis. Instead, chromosome (YAC) transfer is selected in rare segregants ("YACductants") from a yeast mating that is rendered incomplete in most cell pairings by the presence of a kar1 (karyogamy-deficient) mutation in either parental strain. The Basic Protocol in this unit details the transfer of a YAC clone from yeast strain AB1380 (host to nearly all existing YAC libraries) to YPH925, a strain with nonreverting genetic markers compatible with existing plasmid constructs useful in YAC modification.

Original languageEnglish (US)
Pages (from-to)Unit 5.14
JournalCurrent protocols in human genetics / editorial board, Jonathan L. Haines ... [et al.]
VolumeChapter 5
StatePublished - May 2001

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Fingerprint Dive into the research topics of 'Transfer of YAC clones to new hosts by karyogamy-deficient mating.'. Together they form a unique fingerprint.

Cite this