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RNA isolation from isolatedislets and RT-PCR

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#1 anonymous



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Posted 15 July 2001 - 09:00 PM

Hi friends,I was trying to do RT-PCR in isolated pancreatic islets.I have used the primers for which I got good results in the brain regions. But in the case of iselts I didn't get any bands. The problem won't be with the protocols, but with the RNA degradation while RT-PCR. Can you suggest any method to block the Rnase activity of Enzymes which we use for the RT-PCR. I am Using Human placental Rnase Inhibitor in the PCR mixture. I have tried with different concentrations of RNA then also I didn't get anything.Then I have come to a conclusion that RNA is getting degraded during the reaction. Now I am thinking to increase the concentration of inhibitor. Is it enough or you have any suggestions please help me? I am doing Ph.D in Cochin University, Kerala, India. As I have to submit my thesis by the middle of August I am in tension. So please help to get good results

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