Northern witth oligos
Ferland Louis H.
ferlandl at ERE.UMontreal.CA
Thu Feb 22 01:35:05 EST 1996
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
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
> instead of using cDNA (random primed-labelled) ??
> How is specificity, sensitivity ??
> thanks for any comment,
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
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