Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protocols and Methods Forum
Register Search Today's Posts Mark Forums Read

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


How do you prepare RNase-free solutions for Northern blotting?

How do you prepare RNase-free solutions for Northern blotting? - Protocols and Methods Forum

How do you prepare RNase-free solutions for Northern blotting? - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 11-12-2004, 04:25 PM
Paul Wary
Guest
 
Posts: n/a
Default How do you prepare RNase-free solutions for Northern blotting?



Hi,

How do you prepare RNase-free solutions for Northern blotting? For
example, I want to prepare RNase-free 20X SSC or solutions containing
SDS. I planned on using 0.1 % DEPC to make the solutions RNase-free
but I wonder whether DEPC may react with any of the components. And
since autoclaving is necessary for deactivating DEPC, can the
solutions be autoclaved without degradation of any components?

Paul
Reply With Quote
  #2  
Old 11-13-2004, 01:44 AM
Kyle Legate
Guest
 
Posts: n/a
Default How do you prepare RNase-free solutions for Northern blotting?

Paul Wary wrote:
Anything that can be autoclaved and does not contain amine groups (such as
Tris) can be treated with DEPC and autoclaved. Anything that cannot be
autoclaved (some metal salts, etc) or contains amine groups can be made in
baked glassware with DEPC-treated water and dedicated chemical stocks (ie.
no spatulas, no replacing overweighed amounts).

By the way, if you're only planning on using the 20X SSC for transfer to the
nylon membrane, you don't have to worry about making it RNAse free. The salt
concentration is too high to support RNAse activity.


Reply With Quote
  #3  
Old 11-13-2004, 04:35 PM
Nick Theodorakis
Guest
 
Posts: n/a
Default How do you prepare RNase-free solutions for Northern blotting?

On 12 Nov 2004 08:25:10 -0800, [Only registered users see links. ] (Paul Wary) wrote:


I gave using DEPC many years ago, and my RNA looks fine. I do use very
good water (18 mega-ohm, low TOC, etc., as in MilliQ or equivalent),
however.

Nick

--
Nick Theodorakis
[Only registered users see links. ]
contact form:
[Only registered users see links. ]
Reply With Quote
  #4  
Old 11-14-2004, 08:58 PM
fabio
Guest
 
Posts: n/a
Default How do you prepare RNase-free solutions for Northern blotting?

Paul Wary wrote:

Besides what said by Kyle Legate that is certaintily right (like the
reply to a similar my question of 01 november) I can add thar I have
recently read on the Ambion catalog of an alternative reagent to the
DEPC, that can be used also with Tris (RNAsecure...) and of a solution
that decontaminate the electrodes of pH meter (Elecrozap) (that usualy
I don't know how to decontaminate).
Anyway I never used this products so i can't say if they are realy good
and advantageous or not.
Bye.

Fabio
Reply With Quote
Reply

Tags
blotting , northern , prepare , rnasefree , solutions


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
>>> SEGA GAMES <<< Tonia Martinez Forum Physik 0 08-02-2009 02:35 PM


All times are GMT. The time now is 01:40 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.12865 seconds with 16 queries