Like so many on here, I have been struggling with gel extraction kits in my attempts to isolate a ~40 kb phage genome from a mixed sample.
I decided to attempt the gel extraction method detailed by Gustavo here:
https://www.researchgate.net/post/Can_anyone_help_me_to_find_a_protocol_for_gel_extraction_without_using_kits
After the preciptiation with 2.5 vol absolute ethanol and 1/10 vol 5M NaCl I saw what looked like really nice pellets. I then washed the pellet with 70% ethanol and allowed the pellets to air dry until they were just translucent (5-10 min). I then attempted to resuspend the pellets in water, but they would not go into solution. I tried leaving the samples at 55 C for a while, but that didn't work. Finally I gave in and left them at 4 C overnight, but the pellets were still just as large and evident as they were yesterday.
Any ideas whats going on here? Could it be residual agar? It shouldn't be protein as the starting sample run on the gel was from a plasmid maxiprep and was decently pure (104 ng/ul, 260/280 = 1.84, 260/230 = 1.97).
Thanks for any help.