Hi researchers,
I'm using a PCR mycoplasma detection protocol which is found online and it works well for me. Link as follow:
https://bitesizebio.com/23682/homemade-pcr-test-for-mycoplasma-contamination/
I have a problem with this protocol. My positive sample will give a 500bp product, same as the protocol mentions. But besides that, for some samples, I can also get a 100bp product. I'm not sure what it is because I did a blast to check all the primer pairs and their products were really around 500bp, no 100bp one.
Anyone use this protocol before and know what's the 100bp product is?
Thanks!
Sorry I forgot to copy the gel image but the 100bp product is not primer dimmers because the negative control doesnt have this band.
is it coming from the cell line that you are growing and not from the mycoplasma infection. Do the primers blast positive against the cell line dna perhaps ?
Dear Paul,
I test all my cells in culture, for this 100bp sample, not sure it's mycoplasma positive or not. I think it's not cell line DNA cause when I choose human database to blast, there is nothing.
Dear Shuaichen Liu,
Were you able to solve the problem of the extra band?
I'm also curious: Can you freeze the supernatant of your cultures for later processing? I have some negative and positive controls I'd like to save for later testing.
Thanks,
Danuta
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