Escherichia coli phage Phi X174|273419 |BNCC

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Escherichia coli bacteriophage Phi X174

Subculture procedure	phage culture: ① Host bacterial preparation: pick a single colony of host bacteria from a Agar plate, dissolve it in 40-50 mL nutrient broth culture medium, shake at 37 ℃, 160-180 rpm until OD600 is around 0.4; ② Transfer: Remove the phage Frozen vial, rapidly melt it in a 37 ℃ water bath, and transfer all of it into the host bacterial solution mentioned above; ③ Cultivation: Place the triangular flask at 37 ℃ and shake at 160-180rpm overnight for cultivation; ④ Filtration: Centrifuge the phage culture at 4000g for 10 minutes, take the supernatant and collect the filtrate through a 0.22 μ m sterile filter. The filtrate can be stored at 4 ℃
Growth conditions	37 ℃; 18-24h; aerobic;
Storage conditions	-80 ℃
Safety level	2
Sharing mode	Public welfare sharing

Escherichia coli bacteriophage Phi X174

Storage conditions: 2~8 ℃

No. 273419

Product format: freeze dried,200ul

Validity period: 6 years

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

Receiving notice: if any abnormality is found on the day of receiving goods, please contact customer service within 24 hours in time. if it is overdue, it will be deemed as good receipt. Dry powder shall not be retained after one use. please activate it strictly according to this instruction. otherwise, replacement service will not be provided for abnormal strain and inactivation.

Host: Escherichia coli BNCC332831

Host culture conditions:

37 ℃, aerobic, nutrient agar medium, 18-24h. Nutritional agar medium: beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min.

Phage recovery steps:

① Before opening the ampoule tube, prepare a host bacteria solution containing 5~10mL of culture for 18-24h;

② Open it in the safety cabinet, burn the top with an alcohol lamp, quickly drop sterile water to break it, and then crush it with tweezers;

③ Absorb 0.5mL of liquid culture medium into a freeze-drying tube, fully dissolve and then into a host bacteria solution test tube, and mix well;

④ Place the liquid test tube under the above culture conditions and culture for 24 hours.

⑤ 4000 r/min minutes, centrifuge for 15min, take supernatant.

Quality inspection record: activate according to the above activation steps, take the supernatant, drop it on the host plate for 18-24h, and culture at 37 ℃ for 24h. the plaque is obvious. the result is shown in the figure:

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