I came across conflicting protocols online about how lentivirus should be thawed properly before transduction. I saw either to thaw it rapidly at 37°C then keep it on ice, or to thaw it slowly on ice. What would you recommend? Does it depend on the virus buffer? My lentiviral library is in low-protein Gibco OptiPRO SFM buffer, so I guess I should thaw it slowly on ice.
I don't know particularly for the viral libraries, but in general the molecular biological libraries are recommended to thaw slowly on ice. This causes least damage to the structure, theoretically.
It would be great to know is there any difference found in expression of the gene following respective methods i.e., thawing at 37C and 4 degreesC. After your results, please write about your observation.
To my understanding, I don't think there should be any possible difference. Thanks.
Hi Valentin,
In my lab, we leave to thaw the aliquots of vectors at RT before use, but we use them fastly after thawing. If you have many different aliquots, my advice is to keep them at 4°C before use. Indeed, I think thawing at 37°C can induce a thermal shock which is not good. With our process of use, we measured a very high efficiency with standard vectors: 1E9-1E10TU/mL. Furthermore, we did several tests of temperature: 72h at 4°C: no significative drop of efficiency - 72h at RT: drop of efficiency: 1 log.
Hope this will help you
Regards
Nicolas
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus