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Where can I find information on your yeast knockout collections?
Yeast Knockout useful information:
BY4730 MATa leu2D0 met15D0 ura3D0
BY4739 MATalpha leu2D0 lys2D0 ura3D0
BY4741 MATa his3D1 leu2D0 met15D0 ura3D0
BY4742 MATalpha his3D1 leu2D0 lys2D0 ura3D0
BY4743 MATa/alpha his3D1/his3D1 leu2D0 /leu2D0 lys2D0/LYS2 MET15/met15D0 ura3D0 /ura3D0 (4741/4742)
**homozygous diploids are in the BY4743 background unless 4730/4739 is indicated
For an explanation of deletion strain confirmation:
For each ORF, two independent deletion mutants must be produced. This can be done in either of three ways:
1. Transformation of haploids. The diploids are then obtained by mating.
2. Transformation of diploids. The haploids and homozygous diploids are obtained by sporulation and mating
3. Transformation of one of the haploid strains and the diploid strain. The other haploid strain is obtained from the diploid by sporulation and the homozygous diploid is made by mating of the two haploids. The genotypes of the a and alpha segregants have to match those of BY4741 and BY4742, respectively.
A-KanB and KanC-D tests must be positive with the deletion mutant and negative with wild-type. If one of these reactions doesn't work, the A-D* test can substitute.
A-B and C-D tests: Both must be negative with the deletion strain and positive with wild-type.
A-B, C-D, A-KanB and KanC-D PCR products must be present. An A-D* test can substitute if either the A-KanB or KanC-D PCR doesn't work.* Because the sizes of the A-D bands can be similar in wild-type and mutant strains, it can be helpful to digest part of the A-D reaction with HindIII to test for the HindIII site in the KanMX4 cassette
A-B and C-D products must be present in wild-type but not in the mutant. For at least one deletion-specific primer pair, either an A-KanB, KanC-D, or an A-D band of the expected size must be present.
Must be met+lys+, streaked to single colonies, and tested for both mating and sporulation.
Link to list of deletion primers on SGDP: