I am not getting enough concentration of my amplified gene in PCR reaction for cloning purpose. I am trying to increase concentration of my amplified gene by doing rePCR of my PCR product by using PCR product as a template, but this doesn't work. I would like to know is PCR of PCR product would work or not? also how to increase the concentration of gene of interest by PCR as I got thin band of my gene of interest which may or may not be sharp and also got other bands. So I use gel extraction kit method to purify my gene of interest but there is too much loss of product and some times I do not get visible product and enough concentration. Also PCR of such purified product again gives multiple bands which I am not expecting so it is difficult to me to increase concentration of my gene of interest for cloning.
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if you are using boiling method please use commerical kits. Sometimes it is helpful to get DNA. But if you already used it, you should check primers, i had same problem, i had to add one more primer to see band profile.