Technique / Molecular Biology / Cloning library construction and screening
LiAc/SS-DNA/PEG Transformation
LiAc/SS-DNA/PEG Transformation (archive)
High efficiency tranformation protocol from Gietz lab.
Last update 13-Feb-2001, Rating Good of 0 votes.
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By Lindus Conlan
on 05-Mar-2001
There may be a few reasons. Firstly, it is very important to have healthy yeast, pick a single colony from a plate no more than one month old. On the day of growth, the OD600 MUST be started at 0.1-0.2 at then harvested between 0.5-0.6. Other factors include the quality and concentration of PEG-3300 (we use Sigma), don't autoclave your glucose..filter sterilise, don't use DMSO to shock the cells. I would recommend the protocol listed on the Herskowitz Lab website. I found this gave higher transformation efficiencies than the Geitz lab's. |
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By ZJ
on 13-Feb-2001
I have done yeast transformation for some times by Liac and electro. Unfortunately, No any transformants obtained. What is the problems? Any advice for me to set up the transfromation process? |
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