R5421 Yeast Competent Cells

YCC-020

R5421 Yeast Competent Cells

100 μl*50

Saccharomyces cerevisiae strain R5421 is a K+-deficient strain, also known as CY162. The MATα type is mostly used in K+ transporter protein identification assays, but also in K+ channel or sodium-potassium ion pump identification assays. The strain can grow normally in a medium containing 100 mM KCl and grow slowly in a medium containing 5-10 mM KCl. R5421(CY162) cells stopped growing when the KCl concentration in the medium was lower than 0.5 mM.

MATα ura3-52 leu2 trk1Δ his3Δ200 his4-15 trk2Δ1::pCK64

ura3, leu2

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