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Home | Resources | FAQs
Where can I get an explanation of the Yeast Tet-promoters Hughes Collection (yTHC) primers?
Explanation of various primers listed for yTHC collection:
"List of primers" has all the primers (you have to look up by gene).
The next file, "Hughes Lab TET collection strains", says which were confirmed with "inside" or "outside" primers. It was not recorded which outside primer was used.
What each of the primers are:
UP_Primer, Dwn_primer (used to amplify the kanamycin cassette)
UP_Extenders, Dwn_Extenders (used to amplify the amplified kanamycin cassette in order to get longer homology arms - not necessary but might help)
Up_Conf, Down_Conf (these are flanking primers used for confirmation)
"Inside" means confirmation with one of the flanking primers vs. an internal primer in the kanamycin cassette. "Outside" means PCR'd with the two confirmation primers, which is better (because it verifies that the original allele is gone, and that it's been replaced with the tet allele) but that did not work in many cases, even though the "inside" PCR worked and the strain had a growth defect indicating that the wildtype allele was gone.