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.