Northern witth oligos

Ferland Louis H. ferlandl at ERE.UMontreal.CA
Thu Feb 22 01:35:05 EST 1996


SEHuang,

Yes, we've done quite a bit and it works following the principles you 
know: The lower the Hyb temperature (forced down by shortness and A/T 
content of the oligo), the less good specificity; The lower the specific 
activity (only one labelled atom, but gamma-P32-ATP comes in higher spec. 
act.), the less good limit of detection.

In our hands, 30-mers labelled with 6000 Ci/mmol gamma-P32-ATP always 
gave satisfactory bands, but I never tried this on a rare message. (it is 
clear that the limit of detection is lesser than with 
homogeneously-labelled probes, especially riboprobes.)

If your target has a medium or high abundance, it shouldn't give you any 
problem.

Louis



On 21 Feb 1996, SEHuang wrote:

> Date: 21 Feb 1996 08:45:04 -0500
> From: SEHuang <sehuang at aol.com>
> To: methods at net.bio.net
> Subject: Northern witth oligos
> 
> Has anybody tried to hybridize northern blots with oligos (eg T4 kinase
> labelled)
> instead of using cDNA (random primed-labelled) ??
> 
> How is specificity, sensitivity ??
> 
> thanks for any comment,
> 
> S.E.H.
> 

Dr. Louis H. Ferland
Centre de Recherche, Hotel-Dieu de Montreal
Dept de Nutrition, Universite de Montreal
Phone: (514) 843-2757     FAX: (514) 843-2719




More information about the Methods mailing list