We routinely use a kit from Roche in our lab: still, you have to take extra care with RNAse contamination, but the protocol is easy, the kit solutions are already RNAse free and it takes about 1 hour to get a nice amount of RNA.
On Mon, 17 Jan 2011 11:01:48 -0500, Jayakumar, R wrote > 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:methods- > [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 Rosario Díaz-González, PhD Instituto de Parasitologia y Biomedicina "Lopez-Neyra" Avda. Conocimiento S/N Parque Tecnologico Ciencias de la Salud 18100 Armilla - GRANADA Spain Tlf: 958181621 Fax: 958181632 http://www.ipb.csic.es -- Open WebMail Project (http://openwebmail.org) _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
