Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

RNA Buffer Extraction

sodium citrate N-lauryl sarcosine RNA extraction

  • Please log in to reply
3 replies to this topic

#1 Flopilus

Flopilus

    member

  • Active Members
  • Pip
  • 17 posts
1
Neutral

Posted 11 January 2012 - 09:53 AM

I'm preparing an extraction buffer for RNA which has 4 M guanidine thiocyanate, 100 mM Tris-HCl, pH 8.0, 25 mM sodium citrate pH 8.0 and 0.5% N-lauryl sarcosine.
I'd like to know how to bring the sodium citrate solution to that pH, only by dissolving the drug in water?
and, can i autoclave it before adding it to the buffer solution?

Besides, all I have in my lab is N-lauryl sarcosine sodium salt, which I believe is used in RNA/DNA extraction protocols, but I'm not sure to use it likewise...

And finally, can I autoclave the finished buffer?

Thanks in advance!

#2 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,758 posts
130
Excellent

Posted 12 January 2012 - 08:31 AM

never autoclave a solution that contains a detergent.

add all the components (except detergent), adjust the pH, (add the detergent) then bring to final volume .

filter sterilize.

Edited by mdfenko, 12 January 2012 - 08:33 AM.

talent does what it can
genius does what it must
i do what i get paid to do

#3 Flopilus

Flopilus

    member

  • Active Members
  • Pip
  • 17 posts
1
Neutral

Posted 12 January 2012 - 10:11 AM

I've autoclave another buffer with CTAB before and it worked perfectly, don't know why you say I can't...

What I want to know is how to bring to pH 8 the sodium citrate solution (it has a pH of around 9.5 at 100mM), and if I can autoclave it then.

Thanks

#4 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,758 posts
130
Excellent

Posted 13 January 2012 - 06:21 AM

many detergents will solidify when heated. it's just better to avoid if not sure.

you can adjust the citrate the same way you adjust any buffer salt, you can use an acid compatible with your solution or equimolar citric acid to reduce the pH (if separate from the final solution).

citrate can be autoclaved.
talent does what it can
genius does what it must
i do what i get paid to do





Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.