By extracting with CTAB method usually we think that after centrifugation if good quality of Pallet is obtained than there must be presence of DNA. If no DNA is detected on 0.8% gel than what could be reasons.
Hi !
Can you precise the type of plant material you are working on and the whole protocol of the extraction ?
Sincerely
Have you treated the solution by RNAse before precipitation. If yes, check your RNAse quality. The pellet obtained may be of RNA.
You would not expect to see genomic DNA from plants on an agarose gel. The concentration is usually too low to see. Test out your concentration with a Qubit or nano drop or spectrophotometer.
During CTAB method of DNA extraction, oftentimes the following can happen.
1. If you over-dried your pellet after washing phase it cannot be fully mixed with water so that it can reduce the concentration.
2. That DNA pellet may have a kind of polyssaccrides, phenols and other compounds that you had better check the concentration on nano-drop spectrophotometer machine before you go for electrophoresis.
3. The agarose you used may be small, make it 1%.
4. Use RNase during your extraction.
5. If you use sample stored under -80 for long time, degradation can happen.
6. The time and rpm of centrifuge can be matters most. Therefore, use the good protocol most paper published on. Hope you can get the good result. If it become worse use kit for extraction.
Glucides and polyphenols can interfere with DNA extraction. In some case, you will have to try multiple approaches to get the best results : like CTAB + chloroform/IAA, or triazol, or a kit.
Sincerely
Dear Dr Faurie I am thankful for your concern regarding me. I am dealing with Salix species (Willow) Protocol is 1) Crush 1gm leaves(Immature) in Liquid nitrogen and add 7ml DEB(EDTA+Tris-Cl+CTAB+NACL+mercapoethanol+PVP) incubate for 1hr at 65 degree celcius 2) Add 7ml PCI(25:24:1) and centrifuge 7800rpm for 20 minutes 3) extract supernatant and add equal volume of CI(24:1) and centrifuge as above again 4)add 1/10 Sodium acetate +2/3rd chilled Isopropanol and keep over night at -20 degre celcius 5) Centrifuge as above and pipeett out above portion and add 70% alcohol keep palletes for drying. 6) add 1xTE buffer and dissolve the pallet. 8) RNAs treatment at 37 degree Celcius and then check on gel.I am uploading one image(moblie).
With Regards
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