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
- - - - -

Antisense probe as negative control for in situ hybridization

in situ hybridization experimental design controls probe design

  • Please log in to reply
1 reply to this topic

#1 hogthehedge

hogthehedge

    member

  • Active Members
  • Pip
  • 27 posts
0
Neutral

Posted 05 August 2014 - 01:42 PM

Hi guys! So I'm doing some in situ hybridization experiments, and I want to design a better negative control, a labelled probe that shouldn't bind to the mRNA transcribed from my gene of interest.

I'm not trained as a molecular biologist, but I am keen to learn these things! So to hybridize to the mRNA of my gene, I design primers that are going to amplify a bit of DNA that is complementary to it, and then in vitro transcribe RNA that is complementary to that, and so it binds to the mRNAs of my gene, yeah?

How would I go about designing an anti-sense probe? What would this bind to? What should I amplify off of (i.e. what should the primers anneal to?)?

Any help or clues would be much appreciated. Gracias!


Edited by hogthehedge, 05 August 2014 - 01:42 PM.


#2 hogthehedge

hogthehedge

    member

  • Active Members
  • Pip
  • 27 posts
0
Neutral

Posted 15 August 2014 - 09:06 AM

I managed! It was the sense probe I was talking about, not the anti-sense probe. It works!! :)







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

©1999-2013 Protocol Online, All rights reserved.