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I'm Jane. I'm working in Vietnam
My interests was molecular biology of bacteria (PCR, Realtime PCR, ...)
I have just received one primer and I myself setup the PCR protocol. The PCR reaction failed and I don't know why? I used the graient temperature, touchdown PCR and the result was also failure. I'am going to use betain (Quigen) and DMSO to setup again the PCR reaction. But I don't know which forms of DMSO that I need.
Thanks so much if everybody help me!
Can you give us your pcr protocol? don't worry we do not require your sequence, if you can disclose then would be a plus. However few information is very vital such as what is the amount of forward primer and reverse primer did you use, their Tm, amoun of MgCl2, Taq and amount of DNA loaded with its concentration.
Generally if no band is seen I usually give the most lowest stringency parameter for PCR reagent and try. Sometimes it is because your DNA template may contain inhibitor. In short try to supply us with all the information you are able to and you can leave out the confidential detail.
Hope to hear from you soonest.