HIV RT PCR KIT - (Feb/02/2006 )
Can any one suggest me a kit for doing HIV detection by RT PCR on ABI 7500 model.I realise kits from AME Biosciences are not available,Artus and abbot not compatable for 7500.Or please suggest me a protocol for doing the same.I need to decide the sequece for designing probes and primers.
What is it you exactly need? Enzymes for RT and PCR on the HIV genome? Or primers and so on?
In my experience, most RT enzymes commercially available are capable of amplifying regions of the HIV genome (starting from RNA). I'm currently working with superscript III one step high fidelity from invitrogen because my PCR product is quite long (3,6 kb) and the RNA has high secondary structure (envelope gene) so I perform my RT at 55°C, which is working fine.
But, other people have used (succesfully) qiagen one step, transcriptor and expand RT from Roche (that is, in this lab). If you search pubmed, a lot of different labs will use different RT enzymes and different DNA polymerases as well, so it's up to you how much you are willing to spend and how high you want your fidelity.