I extracted genomic DNA from macrofungi with CTAB method. In the chloroform step, I saw a yellow layer on the top layer (see pictures, I showed it with number 3). What is it the reason for this? I repeated this chloroform step but the result has not changed. What do you suggest for me?
Thanks..
Hi, I can be wrong, but it's probably some contamination from polysaccharides left from the phenol step. Maybe another chloro step would help..It can also be oxidised remaining phenol from the previous step in case you picked up by accident during pipetting.
Cheers!
- I agree with the idea of the last commentator
- However I would first try a second Trizol/Phenol extraction before proceeding to a chlorform clean up step
It is due to the hydroxyquinoline added to saturate phenol or as said simply you can say that it is due to the remaining phenol. I saturate crystalline phenol and see the same colour. The lower white part is the proteins and other cell debris, so it it not an issue in DNA isolation.
If I remember correct (I made it 9 years ago), you need to take the yellow part and more carefully do not touch the brown part.
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