I used Trizol to isolate RNA and then the DNAzol to isolate DNA from the interphase as suggested by the protocol. But the pellet of DNA is so hard that it is difficult to resuspend. Why should that be the case? Why should DNA be difficult to dissolve? The protocol suggests that the pellet be resuspended in 8mM NaOH to help dissolution, but this does not seem to help. This DNA is devoid of proteins and RNA. And I did not over-dry it.
I've just asked the same question above, but you have problems with next step, that we didn't do yet. We used self-made reagents, but it appears, that with Invitrogen DNAzol there are some difficulties also.
Hello Olga,
I resuspended the pellet in 8mM NaOH, but of course, as I mentioned, it did not dissolve. So, I left it overnight at 4 degrees and next day, I took the supernatant after spinning the vial, so that I do not take the pellet. I did get DNA and I would say it was good enough for PCR amplification.
I had neutralized the 8mM NaOH with HEPES (free acid).
I was planning to do some normal phenol extraction, but I did not try that once I had PCR working.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- DNA