hi,
i did a qPCR to quantify HIV integration using TZM.bl. i did a PCR targeting gag gene in genomic DNA. i can't find an explanation why TZM.bl wild type came out highly positive.
I'm missing smething?
thank you
It depends on what was your target. TZM-bl contain both luciferase and GFP under the control of the HIV promoter (LTR), so the LTR is there. Otherwise, there are some publications showing TZM-bl are infected with MLV. Finally, have you tried any other non-infected cell? Depending on the PCR assay, it might involve a pre-amplification step that can yield false positives.
Thank tou for your reaponse. As i mentioned i targeted gag. The aplicon is correct as i run a gell and the band is of the proper aize and similar to the one obtained by amplifying a gag containing plasmid.
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