Direct retransformation of yeast with plasmid DNA isolated from single yeast colonies using rolling circle amplification

Biotechniques. 2003 Oct;35(4):774-6, 778-9. doi: 10.2144/03354st08.

Abstract

We have efficiently amplified plasmid DNA from single yeast colonies using rolling circle amplification (RCA). The amplified DNA can be directly used for restriction digestion, DNA sequencing, or yeast transformation. The RCA-based high-fidelity amplification would be useful for plasmid manipulation in a variety of yeast-based systems, particularly for high-throughput analyses.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Technical Report
  • Validation Study

MeSH terms

  • Coculture Techniques / methods
  • DNA, Fungal / genetics*
  • DNA, Fungal / isolation & purification*
  • Genetic Engineering / methods*
  • Nucleic Acid Amplification Techniques / methods*
  • Plasmids / genetics*
  • Plasmids / isolation & purification*
  • Restriction Mapping / methods*
  • Transformation, Genetic*

Substances

  • DNA, Fungal