I use acrylamide gel with 8.5% concentration with 50ml volume but I think it takes too long waiting them forming unto gel, about more than 8 hours, even overnight. In contrast, my friend also use the same gel concentration with 10ml volume and it only takes 10 minutes until it becomes gel. I wonder why, because I need the gel to form quickly.
the most likely reasons for slow setting are not using fresh or enough ammonium persulphate or other old reagents or allowing too much oxygen ( air ) access to the gel.Try adding ore APS and covering the gel with cling film.Set up 2 tube with your aps and your friends aps and cap them and see how they set ( just a couple of ml not a full gradient)
To my experience and in agreement what Paul Rutland says, its either a problem with ammonium persulfate (if older than a few days or too low in concentration), with the TEMED or with the acrylamide itself (too low in concentration, too old, …). Oxygen slows down polymerization as well. In case you have spun your acrylamid mixture, de-oxygenation under low vacuum might be helpful.
Hi Juli, I suggest you that increasing the concentration of APS and TEMED. Also, make sure you mix all components well before and after adding APS and TEMED. Hope that would help. GOOD luck.
Raise the concentration of Ammonium persulphate as well as TEMED and use fresh ones. Keep the gel in airtight condition.
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