I am having trouble resuspending purified human genomic DNA. The genomic DNA is purified using phenol:chlorofom from ~ 1 mL of blood. After ethanol precipitation, I normally see a large pellet which is then resuspended in 150 uL of TE (10 mM Tris pH 8.0, 1 mM EDTA).
But the resulting DNA solution is extremely sticky. There are the options I have tried:
1) Incubation at 37C for 1 hr to overnight
2) Incubation at 50C for 1 hr to overnight
3) Incubation at 65C for 1 hr to overnight
The 65C incubations for up to 3 hours do make the DNA solution less sticky, but in the mean times, cause increasing DNA smearing when running on agarose gel. After overnight incubation at 65C, the DNA solution is no longer sticky but shows extensive smearing on gel, indicating DNA degredation/fragmentation at high temperature.
I have also tried resuspending the DNA in 8 mM NaOH, then neutralise it with Hepes, but this doesn't help either.
I am thinking to try silica/glassmilk beads based methodology. Will genomic DNA captured in silica beads be easier to elute and resuspended without the "sticky" issue.