as viruses are notorious for being prone to mutation at high rates its very difficult to clone them though general cloning procedures and cells so in order to solve this problem need specific cell lines
Dear B R Siddharth
you can try the stbl3
https://www.thermofisher.com/order/catalog/product/C737303
e.coli strain that is designed for the lentivirus cloning.
i used it some years ago for this purpose and it works quite well.
best
Manuele
Hi,
My lab used to use Stbl3 to clone engineered virus-like particles (eVLPs) and to clone several plasmids to avoid DNA recombination as well as for high efficiency. Regarding the flaviviruses, there is a paper also used this bacteria for propagation..."https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10629665/". Additionally, you can refer to another paper using competent E. coli strain C4 "Article Successful Propagation of Flavivirus Infectious cDNAs by a N...
".I hope this information is useful for you.
Good luck!
Phan Thi Lam Hong thankyou very much it will be really helpful if it works as i have tried many times to clone flavivirus but i am facing lot of difficulties hope this helps looking forward for good suggestion from you.
Manuele Martinelli thank you very much. just another curious question can we make more competent cells using these cells from thermo in our lab or there are limitations?
Dear B R Siddharth
sincerelly i ever used the commercial cells since the number of lentiviral clones that we produced were limited, but since they are providing it as chemically competent i do not see any reason why you cannot expand and produce your chemically competent stock. Probably you will obtain cells with lower level of competency (more close to 10^6 cfu/ug than the 10^8cfu/ug and is it possible that you have to optimize more your ligation fo obtain some colonies during your cloning.
good luck
Manuele
- Badges
- Science topic