Some months ago, we started using TGX Stain-Free gels (cat.# 161-0181 and 161-0185) from BioRad, and we have experienced many and different problems of polymerization. Sometimes we have to cast gels twice or even three times.
Has anyone ever had similar problems with this kind of gels? Has anyone been able to solve them?
Hi Rita,
In our lab, we use the solutions provided by Biorad, separately, not the TGX Stain-Free gels prepared solutions. However, the problems in the polymerization of gels could be due to the APS and/or Temed proportions. As I read, the APS amount in my protocol is the double in comparison with the amount in the protocol of TGX Stain-Free gels (cat 161-0185) for the 1.5mm glass plate. In addition, Temed is 15 times lower than APS in my protocol, in contrast, it's 10 times lower in the TGX Stain-Free gels protocol (Resolver solution). So you can try to follow these proportions if you want to test it, maybe you can obtain best results regarding the gel polymerization:
Resolver solution: Total Volume: 15 ml, APS 10%: 150ul, Temed: 10ul.
Stacker solution: Total Volume: 5 ml, APS 10%: 100ul, Temed: 5ul.
These amounts have been checked for preparation of gels at 12% and 15%, for 1.5mm glass plate.
I hope you solve the problem!!
Good Luck!
Thanks a lot Victor! We may try to user higher amounts of APS 10%. However, we did not feel like to do it so far because we tried to increase TEMED concentration (as the technician of Bio-Rad suggested) and the result was even worse. The polymerization was so fast that it resulted in a layered gel. I will keep you updated.
maybe APS or TEMED has expired. I always prepare fresh APS 10%. I have used biorad twice and there was no problem in polymerization whatsoever. very nice gel compared to home made ones. if it wasnt APS or TEMED problem i recommend contacting the company itself.
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