i am doing SSCP technique for my PCR product, i make many gels and the bands separated well into 2 single stranded bands instead of one BUT now from 2 weeks ago the PCR bands didnt separated at all.
i try to change the polyacrylamide to a new one, change TEMED and always used freshly prepared APS and TRIS buffer but no separation and at the end i found that the current slowed down to 5 and then ZERO milliamp. and found no current passed across the gel.
i change the power supply and the electrodes
i think there is some wrong in the polyacrylamide gel it self but i don't know whats the problem
i will be highly thankful if anyone can help me or give me advice to do
waiting ur cooperation and reply
SSCP bands don't separatedSSCP Problem
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