Dissolving DNA - finding better techniques - (Sep/03/2012 )
You should also run a small amount on a gel - the genomic DNA should show up as a blob in/near the wells, with a smear below it.
Yes that is exactly what I'm thinking with the gel electrophoresis...perhaps a few (5) samples from test 1,2, & 3 to assess differences. I ran dna test 3 today with lowest rate though I got distracted and not sure if I'd put double the protease concentration as planned with exception for the two last samples when I had ran short of the protease/fg2 mix. I used the 20- 200ul pippettor set at 50ul for protease/fg2 mix, so maybe pippettor is not fully calibrated and will need to test this or maybe its the 1000ul pippettor that is wrong, cause I would have probably been short about 60ul! And I'm positive it wasn't me that was wrong as I made up two batches of protease/fg2, thinking I'd messed up the first. I think in future the best way is to make enough for 4 extra samples rather than exact amount of samples to cover myself.
i decided to incubate last step at 65 d C for 40mins and then set at 55 d C (as adviced by qiagen on separate occassion) to be left for next two days, if they are still not dissolved I will finish the extra 20 mins at 65 d C. Posting this reply from iPhone and will make follow up post clearer to understand.
Pipettes always have a bit of error in them, though it is usually residue sticking to the tips that causes the problems, I usually make up 0.5-1 extra reactions for whatever I am doing, just in case.