I have been working is S.cerevisiae for about 2 years. Up until about 8 months ago, I would have successful plasmid transformations with about 30-100 colonies per plate depending on the size of the plasmid being transformed. Now, using the same plasmids and same protocol, my efficiency has skyrocketed.. I am now getting thousands are colonies per plate. I know I should be ecstatic about the increased effciency, but something feels off.
Thanks in advance!
It could be that a new reagent has for some reason had a dramatic effect on transformation. However you might also be seeing contamination from somewhere. Have you determined that all the new transformants are real and have the proper plasmid and are of the proper genotype?
Each of my plasmids have a selectable marker associated with them, so in theory, they should only grow on the plates if they carry the plasmid. I haven't done any sequencing analysis, but it is most definietly yeast growing on the plates.
Did you do all the negative controls so that if you don't add plasmid then you don't get transformants?
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