ATLAS: An advanced PCR-method for routine visualization of telomere length in Saccharomyces cerevisiae

Int J Biol Macromol. 2016 Dec;93(Pt A):1285-1294. doi: 10.1016/j.ijbiomac.2016.09.006. Epub 2016 Sep 16.

Abstract

Measuring telomere length is essential in telomere biology. Southern blot hybridization is the predominant method for measuring telomere length in the genetic model Saccharomyces cerevisiae. We have further developed and refined a telomere PCR approach, which was rarely used previously (mainly in specific telomeric projects), into a robust method allowing direct visualisation of telomere length differences in routine experiments with S. cerevisiae, and showing a strong correlation of results with data obtained by Southern blot hybridization. In this expanded method denoted as ATLAS (A-dvanced T-elomere L-ength A-nalysis in S. cerevisiae), we have introduced: 1) set of new primers annealing with high specificity to telomeric regions on five different chromosomes; 2) new approach for designing reverse telomere primers that is based on the ligation of an adaptor of a fixed size to telomeric ends. ATLAS can be used at the scale of individual assays and high-throughput approaches. This simple, time/cost-effective and reproducible methodology will complement Southern blot hybridization and facilitate further progress in telomere research.

Keywords: PCR; Saccharomyces cerevisiae; Telomere length.

MeSH terms

  • Base Sequence
  • Chromosomes, Fungal / genetics
  • DNA Primers / genetics
  • Polymerase Chain Reaction / methods*
  • Saccharomyces cerevisiae / genetics*
  • Telomere / genetics*
  • Temperature

Substances

  • DNA Primers