Hi, I have PCR products of about 70bp. One strand is biotinylated, because I used biotinylated forward primers. Now, how do I isolate only this strand? Do I denature first, then use beads to capture the ssDNA? If so, how to do that? I am afraid that once I denature the dsDNA with heat, I won't be fast enough to capture the ssDNA before reannealing. Or do I capture the dsDNA first, then heat-denature the dsDNA so that the non-biotin strand will fall off? The problems hereby is, again, reannealing of the DNA. Second, heat-denature may even break the biotin-bead bond, causing everything to be lost when I discard the sup? Help!! Thanks.
PS: I prefer not to use high salt solutions, as the final purified ssDNA should not hv salt if possible. Regards, gw _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
