Hello,
I have been having difficulties with PCR inhibition due to plant compounds and have attempted using BSA to no avail. A colleague of mine mentioned he had used many brands of BSA before and some had worked and some didn't. Are there different types of BSA? If not why would they differ so much by brand? What type of BSA would be best for preventing plant compound inhibition?
Thanks,
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#2
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Posted 16 November 2009 - 06:48 PM
You will need a molecular biology grade BSA to make sure there is not any nucleases, nickases, and proteases present.
You will also need to make sure that it is not an acetylated BSA. Acetylation kills nucleases, nickases, and proteases but also inhibits PCR reactions
You will also need to make sure that it is not an acetylated BSA. Acetylation kills nucleases, nickases, and proteases but also inhibits PCR reactions
#3
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Posted 19 November 2009 - 12:27 PM
I agree. Non acetylated is the BSA you need. From what I understerstand, when using acetylated BSA, the acetyl group can sometimes transfer to the polymerase, thereby inactivating it.
Sigma has very good non acetylated BSA (liquid or powder).
Sigma has very good non acetylated BSA (liquid or powder).
Theory is when we know everything and nothing is working. Practice is when everything is working and nobody knows why. Here, we combine theory and practice. Nothing is working and nobody knows why.
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