BY4741 brewing yeast|340897 |BNCC

Essential Information Certificate Related Products

BY4741 Saccharomyces cerevisiae

Subculture procedure	① Prepare 1-2 of the above plates; ② After disinfecting the surface of the ampoule tube, open it in a safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Take 0.5mL of sterile water and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into a Agar plate at a rate of 200 μ L per piece and apply evenly; ④ Place the petri dish under the above cultivation conditions for cultivation, and once the bacterial strain grows, it can be used.
Growth conditions	30 ℃; 24-48 hours; aerobic
Storage conditions	2-8 ° C
Safety level	1
morphology	Colony diameter 1-2 mm, circular with entire margins, opaque, creamy white on the obverse, convex in the center, smooth surface, moist and viscous texture, Gram-positive (blue-violet), cocci, purity: pure.
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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