quick hybidizations

anat-hd un691cs at genius.embnet.dkfz-heidelberg.de
Wed Dec 11 13:36:39 EST 1996


In article <32AE92E5.7488 at glaxowellcome.com>, kucera~gt at glaxowellcome.com says...

>> I want to know if anyone has a formula similar to Clontech's expresshyb
>> for nucleic acid hybridizations? 

>I have used the following QuickHyb solution (BRL's recipe) 
>for a long time with great success on colony blots and 
>southerns.  I typically do a 1 hr. prehyb followed by 2 hr. 
>hyb.  for colony blots.  Works great!

>I hyb southerns a little longer, >4 hrs.

>QuickHyb Recipe: I usually make up about 100mls then aliquot 
>into 10-12 ml tubes and freeze.
>6x SSC ;  5x Denhardt's Solution  ;0.5% SDS   ;100ug/ml SS DNA 

Sorry to correct you, but colonies contain an afwul lot of target 
DNA, and the southerns that you mention are presumably NOT
genomic southerns, but PCR or restriction frags ?

Depending on WHAT you want to detect, such short hyb times are 
not extraordinary.... However, in the buffer that you describe
a genomic southern would still take at least 16 h....

BTW, leave out the Denhardt's and use 7% SDS: easier
and less background.

So anybody know the recipe of a true quick buffer ?

clemens




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