(by R.Jayakumar from roswellpark.org)
Mon Jan 17 11:01:48 EST 2011
You might be getting RNAse contamination, which is very difficult to eliminate except if you use dedicated workspace for RNA work. You should use clean labcoats, workspace that has been wiped down thoroughly with agents such as RNAseOUT or RNASE-BIOHIT or some other regaent that can remove RNAses. Also, you need to wipe down everything including gloves, pipettemans and use RNAse free tips and tubes, RNASE free water and other reagents. The 70% ethanol that you prepare must be prepared with RNAse free water. Anything that comes into contact with the RNA prep or yourselves SHOULD BE RNAse free. IT takes some practice but eventually you will get the hang of it. I routinely get >2.00 pure RNA preps with TRIZOL.
From: methods-bounces from oat.bio.indiana.edu [mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of poonam gupta
Sent: Monday, January 17, 2011 12:49 AM
To: methods from iubio.bio.indiana.edu
As i am also using trizol reagent for RNA isolation and mRNA purification in
my plant tissue and its gave good result but there are some problems i am
facing that the RNA band are not so much clear and its gave some time
contamination. i am using1mg plant tissue for RNA isolation in 1 ml.
please guide me. hope for a positive reply
Biochemistry & Molecular Biology lab
Deptt. of Bioscience & Biotechnology
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