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