Technique / Molecular Biology / Cloning library construction and screening

LiAc/SS-DNA/PEG Transformation

Write your comment

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. Good luck! LINDUS Rating: Good Reply

	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? Rating: Good Reply