Hi,
I'm making RADseq libraries using fish fin clips that have been stored in a chaotropic buffer (4M Guanadinium thiocyanate + detergent, EDTA, TRIS and B-ME). Ideally, I'd like to homogenize these samples in the same buffer and use a magnetic bead based protocol for gDNA isolation as columns or liquid DNA isolation is difficult to achieve with my sample size whereas I can automate the isolation with beads.
My concern is that by storing the fin clips in the chaotropic buffer, I won't be able to get rid of RNA with an RNase digestion before I isolate gDNA, leaving RNA in my sample.
As far as I see it, I have two options: (1) live with some RNA in my library prep (and hope it's impact is limited). (2) Add an RNase treatment after gDNA isolation (and risk losing a lot of DNA because the RNA compete for bead binding sites).
Has anyone solved a similar conundrum or have some advice?
Thanks!!!
Hi David,
I never tried what you're talking about but I did not ear qualification of your samples. did you test them with some stuff as bioanalyzer...? on the other hand, in omics studies you can have amount of results, filtering by bioinformatics ways or at the bench just will enrich you results. pull out the RNA by a RNase treatment.
fred
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA