I am using LAMP to amplify 16SrRNA for detection of phytoplasma in napier grass. In the past week, the negative control has been appearing positive on agarose gel electrophoresis. I even tried using PCR water as the sample and it still gives bands. What could be the problem and how do you think I can solve it?
There are multiple reason for negative control to be appearing positive.
It is clear indicate some step you are getting contamination with DNA. So you Have to check with PCR water, PCR buffer, dNTPs, Primers and even Taq-DNA Pol. If still contamination is there, you have to check other consumables including PCR Vial and Pipette Tips. Sterilize every thing and store in sterile condition. Similarly check for contamination in water, buffer and buffer tank.
Thanks guys, i worked in sterile conditions and i finally got results.
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