MS2 Escherichia coli bacteriophage MS2

Storage conditions : 2~8 ℃

No. : 358039

Product format :freeze dried, 200ul

Validity : 6 years

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

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. The freeze dried culture shall be used up once and shall not be retained. Handling instructions of suspension liquid is enclosed. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.

Host: BNCC271970 Escherichia coli

Host culture: 37°C, aerobic, nutrient agar medium, 18-24h.

Nutrient agar medium: beef extract 3.0g, peptone 10.0g, NaCl 5.0g, agar 20.0g (liquid medium not included), distilled water 1.0L, pH 7.0. Sterilize at 121°C for 15min.

Phage culture:

①Host bacteria preparation: Pick a single colony of host bacteria from the plate, dissolve it in 40-50mL nutrient broth medium, and shake at 37°C and 160-180 rpm until the OD600 is 0.1-0.4;

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

③ Dissolve freeze-dried powder: suck 0.5mL of nutrient broth and put it into a freeze-dried tube, fully dissolve it and put it into a conical flask of host bacterial liquid;

④Cultivation: The flask was placed at 37°C and cultured overnight on a shaker at 160-180rpm;

⑤ Filtration: After 16-18 h, centrifuge the phage culture at 4000g for 10 minutes, and filter the lysate with a 0.2 µm sterile filter, which can be filtered twice to completely remove the host. The filtrate can be stored at 4°C.

Quality inspection

① Prepare 16-18 hours of cultured host bacterial liquid, take 5 mL and dilute it by about 200 times (5 mL of host bacterial liquid is dissolved in 45 mL of nutrient broth, mixed well, and then 1 mL of diluted solution is dissolved in 19 mL of nutrient broth and mixed well) ;

②Put 2-3mL of the dilution solution into 1-2 nutrient agar plates, rotate the plate, spread the bacterial solution evenly on the plate, suck out the excess bacterial solution, and use it after drying (too much host bacteria will cover or cause phagocytosis Plaque is not obvious);

③ Take 10ul of the filtered phage stock solution and spread it on the above plate, 1-3 pieces can be dispensed, the plate is placed at 37°C, and cultured for 16-18h or 24h;