Im my experience, when i can't grind well my samples (fungi grown in PD broth), the SDS buffer provides more dna in the final of extraction. But, sometimes my pellet have some color, and I think that is because have some carbohydrates from the media and from the mucilage produced by the fungi (Colletotrichum).
I know the CTAB helps to remove carbohydrates for the sample, and I tried to mix the buffers. When I did it, my buffer was WHITE. When I centrifuged to decant the debris, was formed one "solid' phase on top of the buffer; i think that is the CTAB. This solid phase don't appear just when I mix chorophorm before centrifuge.
I used to see PVP (1 or 2%) in CTAB buffers. Can I do it if using SDS buffer?
I used CTAB 2% or SDS 3% buffers. Sometimes, I add CTAB 10% in the chlorophorm step (200uL if using 600-1000uL of extraction buffer, plus 600 uL of CIA).
Thank you!
Michael, let me see if I understood.
I can use both, but I need to add one, and late the other. I'm right?
I use more the SDS buffer, but sometimes I add later one mount of CTAB. I first extrac the DNA with 600 uL of SDS 3%, and after incubation I add 200uL of CTAB 10%.
Is it better to proceed the extraction just with one? Now, I'm thinking that the use of both can cause DNA loss.
thank you.
It is important not to have mycoplasma contamination, we must check the addition of inhibitors (antibiotics, mainly tetracyclines)
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