Dye does not separe in acrylamide gel - (Dec/06/2016 )

Hi all,

hope you have some clue. I prepared a 5% acrylamide gel for DNA separation using s(at least I think!) same reagents as usual: acryl. 29:1, 10% aps, etc. Gel polymerised, but when I loaded the dye (standard) for the prerun to clean the gel, I saw that Bromophenol blue and Xylene cyanol co-migrated and did not separe as usual!

I have no idea what happened, reagents were the same but acrylamide (not prepared by me, but at least it polymerised): do you have some?

tnx!

are you sure that there was sufficient crosslinker in the acrylamide, maybe it was prepared wrong? are you sure you don't normally use 19:1 (that's what i used for sequencing back in the day)?

check the pH of the running buffer, if it's significantly off then migration will be affected.

Hi, crsslinking worked as normal, I just re-prepared buffers and everything worked again. Tnx for the tip