I'm attempting to extract RNA from oral mucosa cells. For this, I use a brush to obtain a sample and put it into RNAlater solution. Afterthat, I extract RNA with a special kit, but the results are inconsistent, with ratios 260/280 and 260/230 low. How I can improve the quality, and also the quantity of the RNA?
One of the problem of using RNAlater is the high content in salts that can remain in the sample (showing a 260/230 ratio below 1.8). Also, a low 260/280 may indicate the presence of other contaminants, as proteins. You can improve the quality of the RNA by re-extracting (methods based on the use of magnetics beads are the best option in this case). Additionally, with low quantity of RNA, you can use "priming" to obtain a good sample of cDNA (use specific primers for your target gene to produce the cDNA, only amplificating the region of your interest).
Best regards.
Without putting your samples in RNAlater (as mucosa sample are too low to extract RNA, besides putting it on RNAlater may dillute sample volume), just extract RNA from freshly mucosa samples by Trizol method.
i have no experience in this issue ,,, but i think they using RNA later to preserve RNA for longer time
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA