Hello,
I want to culture human PBMCs, Isolating a subpopulation of T cells by means of MACS Isolation technique (Positive Selection Strategy), and then investigating mRNA expression level of some molecules using qRT-PCR; But I’m worry about probable unwanted effects of positive selection strategy on mRNA expression level. I have some questions in this regard:
1-How can I diminish this probable interference? I think that using Transcription Inhibitors before MACS isolation might work. Can anyone suggest a Transcription Inhibitor with the exact protocol which I should do?
2- Is there any other way that I can inactivate my cells before MACS isolation, consequently diminishing unwanted effects?
Many thanks,
Maryam
If I were you, and I were as concerned as you sound, I'd try a negative selection strategy instead of treating the cells with even more compounds...
Thanks Noel; but I can't perform Negative selection due to the selected marker.
