Looks like you misunderstood the original post. He wants to get RNA from an alkaline prep. That is not possible.
Hiranya S. Roychowdhury, Ph.D. Associate Professor Health & Public Services NMSU-Dona Ana Community College 575 527 7725 (office) ________________________________________ From: [email protected] [[email protected]] on behalf of Michael L. Sullivan [[email protected]] Sent: Sunday, March 10, 2013 1:12 PM To: Ed Siefker Cc: [email protected] Subject: Re: LiCl precipitation? DNase-free RNase, that is! On Mar 8, 2013, at 9:55 AM, Ed Siefker <[email protected]> wrote: > Spec reading tells me I got about 150ug of nucleic acid, and > I'm sure that's not all plasmid DNA. Running it on a gel > shows a little bit of plasmid DNA, and a large bright splotch > running around 100bp. No genomic DNA is stuck in the > wells. > > I figure that blotch either has to be RNA, or DNA. If my > lysis was harsh enough that it degraded the DNA to 100bp > fragments, I would expect to see the small amount of > plasmid as a smear instead of a well defined band. > > On Thu, Mar 7, 2013 at 9:42 PM, Hiranya Roychowdhury <[email protected]> > wrote: >> How do you know that you got RNA? Alkaline lysis does not give you globs of >> RNA. >> >> >> Hiranya S. Roychowdhury, Ph.D. >> Associate Professor >> Health & Public Services >> NMSU-Dona Ana Community College >> 575 527 7725 (office) >> >> ________________________________________ >> From: [email protected] >> [[email protected]] on behalf of Ed Siefker >> [[email protected]] >> Sent: Thursday, March 07, 2013 2:29 PM >> To: [email protected] >> Subject: LiCl precipitation? >> >> I did an alkaline lysis miniprep and ended up with gobs and gobs of RNA. >> I added an equal volume of 4M LiCl, chilled at -20C for 40 minutes, and >> spun at 16K Gs for 20 minutes. I don't see a pellet. What am I doing wrong? >> >> _______________________________________________ >> Methods mailing list >> [email protected] >> http://www.bio.net/biomail/listinfo/methods >> >> >> >> _______________________________________________ >> Methods mailing list >> [email protected] >> http://www.bio.net/biomail/listinfo/methods >> > > _______________________________________________ > Methods mailing list > [email protected] > http://www.bio.net/biomail/listinfo/methods _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
