Hello, I have problem of preparing stacking gel of tricine , there is no issue of polymerizing the resolving gel but problem is in stacking gel polymerization even keep it 5 to 6 hour. I have prepared all new solution for tricine while AB-3 prepared 2 to 3 times with correct composition, using fresh APS but there is no fruitful result. kindly suggest me what should i do
Blot out the moisture/water from resolving gel after it polymerizes and give a wash of n-butanol. Afterwards add stacking gel solution, insert the comb and keep the gel casting chamber adjacent to fluorescent light.
I hope it works for sure......!!
Usually in these case problem could be with APS or TEMED.
I recommend you to degas all acrylamide and buffer solutions for stacking gel preparation, plus degas MiliQ water and use it for preparation of APS solution. For staking gel try prepare 20% APS.
Could you write please what reagents and from what producers are you using for this gel?
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