BY4741 Saccharomyces cerevisiae|BNCC340897 |BNCC

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BY4741 Saccharomyces cerevisiae

Culture medium	YM medium (YM): yeast extract 3.0g, malt extract 3.0g, glucose 10.0g, peptone 5.0g, agar 20.0g (without liquid medium), distilled water 1.0L,pH 6.2±0.2. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it in the safety cabinet, heat the tip of ampoule in a flame, quickly drop sterile water to creak it, then break it with forceps; (3) draw 0.5mL of liquid culture medium into a freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly; (4) inoculate a plate with 0.2mL of suspension liquid, repeat the step to obtain two plates; ⑤ Put all the liquid test tubes and plates under the above culture conditions for cultivation, and the strains can be used when they grow.
Growth conditions	30 ℃;24-48h; Aerobic;
Storage conditions	2-8 °
application	Scientific research
Sharing mode	Public welfare sharing

BY4741 Strain

Storage conditions: 2-8 ℃

Validity period: 5 years

No. 353719

Product format: Freeze-dried powder

Biosafety level: 1, handle in ultra-clear table or safety cabinet

Usage: competent cells, plasmids, genetic engineering

Note: this product is transported at room temperature. please store it at -20 ℃ after receiving it. If any abnormal situation such as damage is found on the same day, please contact customer service within 24 hours. The Xilin bottle shall be used up once after opening and shall not be retained. Please operate in strict accordance with this instruction, otherwise the replacement service will not be provided if the strain is abnormal and inactivated.

Medium:YM agar medium yeast extract 3.0g, malt extract 3.0g, glucose 10.0g, peptone 5.0g, agar 20.0g (liquid medium not included), distilled water 1.0L, pH 6.2±0.2. Sterilize at 121℃ for 15min.

Growth conditions

Temperature:30°C

Gas environment: Aerobic

Instruction

A. Recovery

1. Prepare 2 pieces of YM plate;

2. In a sterile environment, burn the top with an alcohol lamp, quickly drop sterile water to break it, then break it with tweezers, and then beat it into 0.5mL of liquid culture medium to fully dissolve;

3. Scribe the suspension on the plate, 2 plates, 24-48h to grow a single colony;

B. Competent preparation (for reference)

1.BY4741 Saccharomyces cerevisiae plate, pick a single colony and connect it to a test tube of 3ml YM liquid medium, and culture at 30 ℃ for overnight.

2. Take 1ml of bacterial solution and transfer it to a conical flask containing 50ml YM liquid culture medium, shake culture at 30 ℃ for 2~3h,A600 should be between 0.4~0.5, and the number of cells must be <10⁸CFU/ml.

3. Transfer 1ml of bacterial liquid to a 1.5 ml centrifuge tube with a pipette gun and place it on ice for 10min.

4. Centrifugation for 10min,4000r /min.

5. Pour out the culture solution and use cold 0.1mol/l CaCl₂ 200 µl suspension precipitation, immediately placed on ice for 30min.

6. Low temperature centrifugation for 10min,4000r /min, cells were recovered.

7. Use ice-cold 0.1mol/l CaCl₂ 200 µl suspension cells.

8. Subpackage cells, each 200ul, this cell is competent cells.

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