Dear all,
All samples are cDNA, using reverse transcriptase enzyme to make cDNA from RNA. One group is doing PCR without DMSO, and other group is doing PCR with DMSO.
One is without DMSO, the clear bands are appeared including no reverse transcriptase enzyme during doing cDNA library.
One is with DMSO, multiple bands are appeared.
Samples dont have any problems, I have checked as the same protocol for other 4 different genes, all are okay.
Could you help me to understand the problem happened in this gene?
DMSO can act a bit like urea in that it can denature molecules by disrupting their hydrogen bonding, it also has a slight tendency to autoxidize generating ROS causing ssDNA breaks. These effects apply to proteins also, so if the DMSO concentration is high expect it to impair enzyme function also.
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