I need to a manual protocol for extraction RNA from Agarose.
I don't want to use any kit for it. I noticed that manual protocols are very well than Kit System.
Please help..
Melik
Are you using formaldehyde gel electrophoresis?
http://link.springer.com/protocol/10.1385%2F0-89603-494-1%3A69
(You cannot use the traditional agarose gel as per DNA)
Yes, but it needs to be a modified protocol. Once he confirms his banding, he can cut the gel the way. Although I agreee with Artur that cutting the gel is a messy way. If you are doing down-stream analysis like RT-qPCR the gel mixture may have inhibitory components for the cDNA synthesis and PCR.
"RNA has the tendency to form both secondary and tertiary structures that can Impede its separation by electrophoresis. As such, identical species of RNA exhibiting varying degrees of intramolecular base-pairing, migrate at different rates and result in the smearing of distinct RNA molecules. Consequently, the electrophoresis of RNA needs to be performed under denaturing conditions. Heat denaturing the RNA sample prior to electrophoresis is insufficient, as secondary structures will simply reform unless a denaturing system is used."
http://link.springer.com/protocol/10.1385%2F0-89603-494-1%3A69
firstly, thank you very much for your ansvers, actually, your informations are very usefull for me. I am so busy :( because I could not write anything. I am sorry. I locked for your said things. So My purpose is to study gene expression and I tried some porotocols about pure RNA and I found a protocol as a manual for extraction of RNA so, I have overcomed this proplem and your informations are heplful, Mr Arthur and Mr. Can thank you again.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Biological Techniques