INVSC1 Yeast Competent Cells

YCC-010

INVSC1 Yeast Competent Cells

100 μl*50

INVSc1 strain is a host yeast specialized for recombinant protein expression. It activates the transcription of the GAL1 promoter of the pYES2 plasmid and thus initiates the expression of downstream recombinant proteins. INVSc1 is a syncytial type, and the pYES2 plasmid can be directly transformed into INVSc1 cells by PEG/LiAc. The screening marker for plasmid pYES2 is URA, and it can be screened by an SD-URA plate.

MATa his3Δ1 leu2 trp1-289 ura3-52/MATα his3Δ1 leu2 trp1-289 ura3-52

pYES2

Transformation efficiency >10⁴ cfu/ug DNA as assayed by pYES2 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.

1). Competent cells are best melted on ice, PEG solution will precipitate at low temperatures, please dissolve completely at room temperature before use.
2). The yeast strain is sensitive to high temperatures, and the optimal growth temperature is 27-30℃. Above 31°C, the growth rate and transformation efficiency decreased exponentially.

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