NMY51 Yeast Competent Cells

YCC-014

NMY51 Yeast Competent Cells

100 μl*50

The NMY51 yeast strain can be directly transformed into plasmids for protein interaction validation or screen library assays.

MATahis3200trp1-901leu2-3,112ade2LYS2::(lexAop)4- HIS3ura3::(lexAop)8-lacZade2::(lexAop) 8-ADE2 G

trp1, leu2-3

HIS3, ADE2, lacZ

The first step is a selective growth screen by nutritionally deficient reporter genes (HIS3, ADE2), and the further step is a quantitative or semi-quantitative screen by the LacZ reporter gene for β-galactose analysis of chromogenicity. Three independent reporter genes, regulated by different promoters, reduce the chance of false positives. This system can use four Bait plasmids pBT3-N, pBT3- C, pBT3-SUC, pBT3-STE, all with the screening marker LEU; and four Prey plasmids pPR3-C, pPR3-N, pPR3-SUC, pPR3-STE, all with the screening marker TRP.

Transformation efficiency >10⁴ cfu/ug DNA as assayed by pGADT7 plasmid.

1). Take a sterile 1.5 ml EP tube, sequentially add 1-3 μg of pre-cooled target plasmid, 10 μl of Carrier DNA (95-100 ℃ for 5 min in a rapid ice bath, repeat once), and 100 μl of competent cells melted on ice, 500 μl of PEG/LiAc, and gently turn and mix 6-8 times.
2). 30 ℃ water bath for 30 min, every 10 min gently turn over and mix 6-8 times.
3). Add 20 μl of dimethyl sulfoxide to each branch (used to improve the conversion efficiency, can not be done).
4). 42 ℃ water bath for 15 min, every 5 min gently turn over and mix 6-8 times.
5).12,000 rpm instantaneous centrifugation and discard the supernatant, add 1 ml of YPD Plus to each cartridge, and resuscitate at 30℃ for 1 h (used to improve the transformation efficiency, can not be done).
6).12,000 rpm instantaneous centrifugation and discard the supernatant, resuspend with 100 μl of 0.9% NaCl, coat the plate, and incubate at 30℃ for 48-96 hours.

Competent cells are best melted on ice, PEG solution will precipitate at low temperatures, please dissolve completely at room temperature before use.

5 μg/μl, 100 μl

5 ml

Competent cells are transported on dry ice and stored at -80°C. Carrier DNA needs to be stored at -20°C. PEG/LiAc needs to be stored at 4°C.

6 months