I am running a gel electrophoresis to check mycoplasma positive samples using PCR, however only positive control shows up.
I tried new master mixes and diluting the samples but in all cases only positive control shows up.
Paul Rutland
Are your samples from cultured cell line or natural samples. How much dna are you adding to the pcr and do you have an OD260/280 ratio for the dna.
How many pcr cycles are you running and are the lanes with your amplimers completely clear or is there a long smear of fluorescence in those lanes.
Are you expecting any of your samples to be positive for mycoplasma
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Sixtus Amarachukwu Okafor
confirm if you actually have DNA present in the sample you loaded on the wells
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Keshav Meghwanshi
Aashka Mehta
you can try using DMSO... And cross check the quality of your template using 260/280 ratio... Further Analyse the primers with your template sequence... Share the gel images, if possible, for better understanding of the issue faced by you.
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