BY4741 Chemically Competent Cell

For the first use of Carrier DNA，boil the tube containing Carrier DNA in boiling water for 5 min，then place it on ice immediately. Store it at -20℃ for standby after use. Thaw Carrier DNA on ice before subsequent use. Re-denature Carrier DNA after 3-5 repeated freeze-thaw cycles. Competent cells are best thawed on ice. The use of high-concentration plasmids for transformation can correspondingly reduce the amount of bacteria ultimately used for plating. Saccharomyces cerevisiae is temperature-sensitive with an optimal growth temperature of 28℃-30℃; temperatures exceeding 31℃ will affect yeast growth. Yeast grows more slowly in deficient medium than in YPDA medium，and the more deficient components contained in the medium，the slower the growth rate. BY4741 is derived from the original Saccharomyces cerevisiae strain S288C，which is a commonly used laboratory strain of MATa mating type. BY4741 is a histidine，leucine，methionine and uracil auxotrophic strain，and the pYES2 plasmid can be directly transformed into BY4741 cells via the PEG/LiAc method. The pYES2 plasmid carries the URA selection marker and can be screened on SD-URA plates. The transformation efficiency of BY4741 competent cells verified with pYES2 plasmid (5857 bp，Amp?) is over 103 cfu/μg DNA. Genotype: MATa his3Δ1 leu2 met15Δ ura3-52