CTAB extraction problem
Posted 08 August 2011 - 08:54 AM
I am not experienced in molecular biology procedures and have encountered some problems in a CTAB DNA extraction in two plant species from the Lauracea family.
When I add the CTAB buffer to the powder, the mixture forms a very viscous substance. The main problem in that after I add the cloroform/isoamylalcohol (24:1) solution and spin at maximum speed, literature says that it is expected that I obtain a three layer fase in the tube; the top fase is the aqueous solution with the DNA and what I should retain (supernatant), followed by a layer with the cloroform/isoamylalcohol and followed by the cell debris at the bottom of the tube.
What I obtain is something quite diferent; the supernatant fase is usually very little and sometimes I donīt obtain any supernatant, followed the cell debris fase and at the bottom of the tube an aqueous layer which I expect should be the cloroform/isoamylalcohol.
Can anybody tell me what can be the problem here. Why donīt I get supernatant and why does the cell debris stay in the middle of twi liquid fases?
Thanks in advance for any help
Posted 08 August 2011 - 04:11 PM
You may need to do your extractions in a bigger volume as well.
Posted 09 August 2011 - 03:23 AM