Chunk #54 — Materials and Methods — Subcloning and strain construction

Source

Replicative age induces mitotic recombination in the ribosomal RNA gene cluster of Saccharomyces cerevisiae.

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yes

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MEP deletion strains for bud6Δ, csm1Δ, lrs4Δ, sir2Δ and sas2Δ were generated by one-step gene disruption and verified by PCR using the following oligonucleotides and DNA templates: BUD6: deletion- Bud6delF/bud6delR, template- pRS306, confirmation- Bud6delcheck/Bud6delchkDN. CSM1: deletion- CSM1A/CSM1D, template- UCC7629-1 genomic DNA, confirmation- CSM1A/CSM1B. LRS4: deletion- LRS4A/LRS4D, template- UCC7598-1 genomic DNA, confirmation- LRS4A/LRS4B. SIR2: deletion- SIR2KO1/SIR2KO2, template- pRS400 [63], confirmation- 5′SIR2/3′SIR2. SAS2: deletion- SAS2KOF/SAS2KOR, template- pRS400 [63], confirmation- SAS2conF/SAS2conR.