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. Jay
-----Original Message----- From: [email protected] [mailto:[email protected]] On Behalf Of poonam gupta Sent: Monday, January 17, 2011 12:49 AM To: [email protected] Subject: Request Respected sir 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 Thanking you -- Poonam Gupta Research Scholor Biochemistry & Molecular Biology lab Deptt. of Bioscience & Biotechnology Banasthali University Banasthali (Rajasthan) 304022 _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
