Up till now I've been attempting to isolate HIV linear DNA through a TOPO-cloning technique. I think I mentioned previously that after coming back from a conference, Una was very excited about moving on to an alternative technique which we had been considering, called the Hirt extraction, which is a method of isolating small nucleic acids within a background of high-weight chromosomal DNA. Theoretically this technique could be applied to HIV as a method of isolating non-integrated species of HIV DNA.
I am now shifting gears to trouble shoot the method, and in addition, we will now start looking at 2 LTR circles, a non-productive non-integrated HIV DNA species that may or may not be indicative of productive/latent infections. There's a lot to work out and I am a little concerned that I won't get as far as I would like before the end of the semester (just a few weeks!). Fortunately I am getting a lot of help from Jenny so hopefully things will go well.
Oh, and there's always the final exam for Cell Biology coming up...
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