Escherichia coli bacteriophage T4

Storage conditions: 2~8 ℃

No. 251275

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 BNCC305635

Host culture:37°C, 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 culture:

① Preparation of host bacteria: single colony of host bacteria is selected from the plate, dissolved in 40-50mL nutrient broth medium, oscillated at 37 ℃ and 160-180 rpm until OD600 is 0.1 - 0.4 ;

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

③ Dissolved freeze-dried powder: absorb 0.5mL of nutrient broth and enter into a freeze-dried tube. after fully dissolving, enter into a triangular flask of host bacterial solution.

④ Culture: the triangular flask was placed at 37 ℃ and incubated at 160-180rpm in a shaker overnight;

⑤ Filtration: centrifuge the phage culture at 4000g for 10 minutes after 16-18h, and use 0.2 & micro;m sterile filter to filter the lysate, which can be filtered twice to completely remove the host, and the filtrate can be in
Preserved at 4 ℃.

Quality inspection

① Prepare host bacteria solution cultured for 16-18h, take 5mL and dilute it about 200 times (dissolve 5mL host bacteria solution into 45mL nutrient broth, mix well, then dissolve 1mL diluent into 19mL nutrient broth and mix well);

② Take 2-3mL of diluent into 1-2 nutrient agar plates, rotate the plates, evenly coat the bacterial liquid on the plates, suck out the excess bacterial liquid, and use it after drying (too many host bacteria will cover or make plaque not obvious);

③ 10ul of filtered phage stock solution is applied to the above plate, 1-3 of which can be applied, the plate is placed at 37 ℃, and cultured for 16-18h or 24h;