Escherichia coli phage T1

Storage conditions: 2~8 ℃
No. : 259107
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. If it is overdue, it is deemed that the bacteria are well. The freeze dried culture shall be used up once and shall not be retained. 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:  Escherichia coli  BNCC305635
Host growth conditions:  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 recovery steps:
① preparation of host bacteria: single colony of host bacteria is selected from the plate, dissolved in 40-50mL nutrient broth medium. Put it in a shaker ( 160-180 rpm) at 37 ℃ until OD600 is 0.1 - 0.4 ;
② Open the freeze dried vial in the biosafety 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 nutrient broth into a freeze dried vial, fully dissolve and transfer the solution to the flask of host bacteria solution;
④ place the flask on a shaker( 160-180rpm ) at 37 ℃ for cultivation overnight;
⑤ after 16-18 hours, the bacteriophage culture is centrifuged at 4000g for 10 minutes, and the lysate is filtered with a 0.2um sterile filter, which could be filtered twice to completely remove the host. Store the filtrate at 4 ℃.
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 spread 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 Take 10ul of filtered bacteriophage stock solution and point coat it on 1-3 of above-mentioned plates. Place the plates at 37 ℃ and culture for 16-18h or 24hours.