RNA Gels - (Jan/18/2008 )

I have been trying to isolated RNA from quail samples stored in RNA later. I am using the RNeasy mini kit from qiagen. When I spec the RNA I get ok yields (1ug/ul) and the ratios are all fine. The big problem is when I run it on a denaturing gel I only get one very bright sharp band (no streaking or smearing). It is at 1.8kb so I think it is the 18s band ... any ideas???? I have tried using fresh quail tissue and I get the exact same thing. I know it is not the kit b/c I have extracted other tissues from different animals as controls and they work perfectly. I think I have tried anything so I'd love so help!!!!
Thanks, Ali

-Ali-M-

Hey I'm only new but... an idea I might have is did you dentaure the RNA - heat it to 70 degrees and then snap cool it first? Maybe the one band is due to the structure.... THIs could be totally off, so sorry if it is!!

-scistudent-

QUOTE (scistudent @ Jan 20 2008, 07:23 AM)

Yeah I heated it and then snapped cooled it before I loaded it on the gel.
Since posting another student has isolated other quail samples and gets the same results... one sharp bright band at 1.8kb.
I have made cDNA from my sampled and used it to PCR a housekeeping gene... thinking I might send sample for sequencing, but I have no idea what is going on with the RNA

-Ali-M-

oh... hmm I dont know then. sorry!

-scistudent-