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!
- - - - -

bacterial RNA isolation from PBS buffer water

Membrane filtration Membrane filter TRIzol Homogenization

  • Please log in to reply
No replies to this topic

#1 Md Jibran Alam

Md Jibran Alam


  • Members
  • Pip
  • 4 posts

Posted 28 November 2014 - 05:38 AM

I want to extract the total  bacterial RNA directly from sample collected in PBS. I cant enrich the bacteria as my research focuses on direct isolation of rna. As bacterial count is less, i also need to filter out the bacteria.


I dont have any bead-beater in my lab. So, I cannot use MOBIO Powerwater kit to isolate rna from bacteria filtered with .22 um membrane filter.


I have TRIzol available in my lab. So I want to follow the steps below. Please tell me if it is okay or suggest modifications what I can do with TRIzol reagent.


Modified Homogenization steps by using TRIzol-


1. separate the bacteria with .22 um membrane filter (1 cm in diameter).

2. put the membrane filter in a microfuge tube.

3. add 750 ul TRIzol reagent inside the tube.

4. crash / homogenize the membrane filter with IQ plus tissue homogenizer.

5. centrifuge for 2 min at 13000 rpm. (???? need more duration??)

6. collect the supernatant from the tube and proceed to the subsequent manufacturer protocol of TRIzol reagent.


Thanks in advance.

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

©1999-2013 Protocol Online, All rights reserved.