McClean:Yeast Transformation: Difference between revisions
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# 50% w/v PEG (mw 3350) (Sigma P3640) | # 50% w/v PEG (mw 3350) (Sigma P3640) | ||
# Sterile H20 | # Sterile H20 | ||
# Single-stranded carrier DNA (2.0 mg/ml in TE buffer pH 8.0) | |||
# Appropriate selective plates (SC-URA, YPD+G418, etc) | # Appropriate selective plates (SC-URA, YPD+G418, etc) | ||
Revision as of 10:25, 20 July 2011
Overview
Our lab's version of the Geitz lithium-acetate transformation method.
Materials
- 1M LiAC
- 100mM LiAC
- 50% w/v PEG (mw 3350) (Sigma P3640)
- Sterile H20
- Single-stranded carrier DNA (2.0 mg/ml in TE buffer pH 8.0)
- Appropriate selective plates (SC-URA, YPD+G418, etc)
Day 1
- Inoculate the strain to transform from a single colony into 5mls of YPD in a test tube. Put on the roller drum at 30°C overnight.
Day 2
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
References
Gietz, R.D. and R.A. Woods. (2002) TRANSFORMATION OF YEAST BY THE Liac/SS CARRIER DNA/PEG METHOD. Methods in Enzymology 350: 87-96.
Contact
- Who has experience with this protocol?
or instead, discuss this protocol.
