Hybridisation oven questions

Klaus.Matthaei at anu.edu.au Klaus.Matthaei at anu.edu.au
Sun Jun 27 18:57:54 EST 1993


>Hi all
>
>We have just bought a hybridisation oven and i was wondering if you all out
>there have any tips on making the most effective use out of it..
>
>ie...
>
>What has been the maximum number of filters used per bottle?

I have used up to 5

>Do you really need to use the mesh provided for placing between filters?

Dunno but always have.

>What kind of hyb solution volume to surface area of filter do people use?

Varies for target and probe i.e DNA-DNA, DNA-RNA etc

>Has anyone altered the hyb soln composition as a result of using these ovens?

No

>Are any brands of filter/membrane better than others in these ovens.?

I use Hybond N+ and alkaline transfer.

>Do people simply use bag hybridisation methods in the ovens?

Why??

>
>..and can you really cook a chicken in one of them :-)
>
>Any help on this would be gratefully appreciated. If i get enough response, Ill
>try to get a summary sorted out for everyone.
>
>Thanks
>
>Nigel Walker
>
>Johns Hopkins, Baltimore, USA


Hybe ovens are great but one word of caution.

GET ONE WITH AN OVERTEMP CONTROL.  
One of my hybes sat at 135 (yes 135)*C overnight.  The bottles are rated to
70*C but luckilly did not explode.

Cheers, Klaus
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Klaus Matthaei
Gene Targeting
The John Curtin School of Medical Research
The Australian National University
E-mail: Klaus.Matthaei at anu.edu.au

"I know you think that you understand what I am saying."
"But what I am saying is not what I mean."
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