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Colony PCR Using Yeast Spheroplasted Cells

Bio-protocol Editor Bio-protocol Editor

Published: Jan 5, 2011 DOI: 10.21769/BioProtoc.10 Views: 14294

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Abstract

The first part of this protocol involves the removal of yeast cell walls using the Zymolase enzyme. The resulting spheroplast cells can then be used as template for PCR. This quick and easy to implement protocol describes how to prepare spheroplasted yeast cells for colony PCR.

Materials and Reagents

  1. Spheroplasted yeast cells
  2. Zymolase 20 T [AMS Biotechnology (Europe)]
  3. Sorbitol
  4. Sodium phosphate
  5. Zymolyase solution (see Recipes)

Equipment

  1. Sterile pipette tips
  2. Standard laboratory PCR machine
  3. Wooden toothpicks

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Molecular Biology > DNA > PCR

Microbiology > Microbial genetics > DNA

Cell Biology > Cell structure > Cell surface

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useful
edit 0 Answer 3 Views Nov 19, 2012