|Culture medium||(Host: Escherichia coli BNCC354694) Nutritional agar medium: beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (without liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min.|
|Growth conditions||37°C;18-24h; Aerobic;|
|Storage conditions||2-8 ℃|
|morphology||Plaque is obvious|
|Sharing mode||Public welfare sharing|
Escherichia bacteriophage Qbeta
Storage conditions: 2~8 ℃
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 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. The bacterial viability will resume after 1-2 generation of recovery and can be used normally. 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 BNCC354694
Host growth:37°C, aerobic, nutrient agar medium, 18-24h. Nutritional agar medium: beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (without liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min.
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 phage culture at 4000g for 10 minutes after 16-18h, and use 0.2 & micro; M sterile filter to filter lysate, which can be filtered twice to completely remove the host, and the filtrate can be stored at 4 ℃.
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;