Hello everyone ,
I'm trying to do transfection by lentiviral vectors and my packaging cell line is HEK293T.
We are going to do calcium phosphate method.I would like to find the best protocol for this method.
The second generation lentiviral vectors are available as follows:
pSPAX
pMD2G
pCDH-insert
Any suggestion or comment in regard to my question will be highly appreciated .
Best,
Niloufar
There are small variations of the standard protocol which in the 293T case most likely make no much difference. So use the one most familiar to you. The viral titer however may vary a lot depending on the lenti-construct..
Dear Joseph ,
Thank you for your answer.
Best
Niloufar
@JosephPapamatheakis
HEK293T cell line is the easiest cell line to transfect. Everything works. As Joseph Papamatheakis have written, the protocol for calcium phosphate transfection has only some small variations. So, just find one and use it. However, with lentiviral transduction later, it is worth to think about additional use of Polybrene, especially if the virus titer will not be very high.
Dear Patrycja ,
Thank you for your consideration.
Best
Niloufar
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