BeNa Culture Collection
Culture medium | EMEM complete medium (containing 2% FBS):98% EMEM + 2% FBS |
Subculture procedure | ① Host cell HEL 299 culture
the frozen cells were recovered to T25 bottle for culture, and the density could reach 80% for 3 days under the culture conditions of culture temperature 37 ℃ and gas environment 5% CO2 + 95% air. at this time, the virus was inoculated. ② Virus inoculation Dissolve BK polyomavirus in a 37°C water bath. After removing the cell culture medium from the HEL 299 at a density of 80%, the residual medium was removed by gently rinsing with PBS for 1-2 times. Inoculate 1ml of virus solution into host cells, adsorb for 1-2 hours at 37°C, 5% CO2 + 95% air, and gently shake the T25 bottle every 15 minutes to make the virus distribution more uniform. ③ Virus culture After adsorption was completed, adsorption was terminated by adding 6-8mL of virus growth medium. Place in an incubator for 14-23 days. Daily observations are required, and virus collection begins when all host cells have developed lesions. ④ Virus fluid collection The cells were repeatedly frozen and thawed twice at harvest, and then the virus was collected with a 0.22 μm pore size filter. |
Growth conditions | Culture temperature 37 ℃; Culture time 14-23 days; Gas environment 5% CO2 + 95% air; |
Safety level | 2 |
morphology | cell degeneration; cell rounding; |
Separation substrate | The strain was isolated in 1970 from the urine of a kidney transplant patient in London. |
application | This reference was developed by Henan engineering center of industrial microbial strain. It is an inactivated particle of BK polyoma virus. It can be used for quality control of BK polyoma virus, development of standard substance, positive control of nucleic acid detection, nucleic acid detection research, etc. |
Sharing mode | Public welfare sharing |