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Joe Horecka and Angela M. Chu 2017. Yeast Colony PCR: It doesn't get any easier than this!. protocols.io https://dx.doi.org/10.17504/protocols.io.gzwbx7eCopy Citation Copied
URL: https://dx.doi.org/DOI:10.17504/protocols.io.gzwbx7e
Authors: Joe Horecka and Angela M. Chu
Group: Genomes to Natural Products Network
Summary: A protocol for yeast colony PCR, starting with intact cells. Can be used to analyze genomic and plasmid DNA, with PCR products up to 2 kb being no problem. It doesn't get easier than this: pick a small amount of cells from a plate into water, lyse cells at 99C for 5 min. Use the lysed cells as template by adding to 1/10th final volume in a PCR. We use this protocol for diagnostics (Phire HS II enzyme) and for Sanger sequencing (Phusion HS II or NEB Q5 enzyme). The main reasons why others might not get this protocol to work could be due to using too many cells and/or not using pure water for the cell lysis. Good luck, and have fun! See the 'BEFORE START' section of this protocol for photographs that show how to pick cells from a colony and make a suspension. See the 'WARNINGS' section for important points and trouble shooting.
Affiliations: Stanford Genome Technology Center
Version: 1
Publication Date: 2017
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