I have a rather simple question. I need to know what the minimum # of
transformed colonies people have used to clone their genes of interest.
I know the figure of 100,000 (Guide to Yeast Genetics and Molecular
Biology) is often used, but can anyone give me the rational behind this
(reference)? I also know of the formula for determining the number of
clones required (N) to isolate a gene with a certain probability (P),
but when I use this formula to calculate (N) with a (P=0.95) for a
library containing 10-15 kb inserts I come with something around only
5,000 clones. Finally, I have read a paper by Jasper Rine, where he
he describes targeted selection of recombinant clones through gene
dosage effects. In this paper he describes the cloning of a Tunicamycin
resistance gene with a multi-copy library with only 4,000 transformants.
I would appreciate answers for both CEN and 2-micron based plasmids.
Thanks for any answers or ideas,
-AL(aplummer at sprint.uccs.edu)