Could you specify the down-stream application? Are you amplifying pro-virus? Typically you don't have to stimulate them to improve DNA yield, stimulation usually is used to change the transcript (mRNA) expression profile. For DNA, you can just have more cells to extract from.

Hi Kejun, thanks for your reply. Yes, I'm interested in getting more cells really, sorry for the misused term. We are indeed interested in amplifying pro-virus for sequencing.

Thanks again!

So, if I understand it correctly, what you plan to do is to stimulate PBMC so that them can be infected by HIV-1. Are you amplifying HIV for detection (infected or not) or HIV genome recovery (amplify whole HIV genome)?

Hey Kejun, I just need to amplify for integrated HIV genome for downstream sequencing.

Typical PBMC stimulation uses PMA-ionomycin method. Detailed protocol you can google it online. If i remember correctly PMA at 50ng/ml and ionomycin at 500ng/ml...

Just curious, are you sequencing the HIV genome for phylogenetic analyses or studying HIV mutation during infection?

Thank you! It's just to see the mutations in different patients during the course of their disease on the first place!

That’s interesting! Are you going to traditional Sanger sequencing or next-gen high throughput sequencing methods?