Abstract
The reverse transcription (RT) reaction is a critical step in HIV-1 life cycle. It is very strongly regulated and the target of several restriction factors (TRIM5α, APOBECs, SAMHD1, etc.). The progress of reverse transcription can be followed by measuring viral DNA by quantitative PCR (qPCR). This method is sensitive enough to allow detection of low amounts of HIV-1 DNA in infected cells and discriminate between several types of reverse transcription intermediates (so called 《early》 and 《late》 RT products, 2 Long Terminal Repeat (LTR) circles, integrated DNA).
Materials and Reagents
Equipment
Procedure
Notes
Acknowledgments
This protocol is adapted from Roesch et al. (2012).
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.