HELP! Desperately need a dot blot method

Gregg Silk greggsilk at aol.com
Tue Jun 30 07:19:06 EST 1998


>I may have a simple solution to your problem.  When I do colony lifts of
>E.coli containing plasmids of interest I use the "Turbo Juice" method. 
>Basically this involves using dry membrane to lift the colonies.  The
>membrane is then put DNA side up on a bit of 3M paper that has been
>soaked with Turbo Juice (simply 2x SSC + 5% SDS) and popped in the
>microwave on high for 3-4 minutes until very hot and dry.  The membrane
>can then be hybridised straight away; no washing or UV linking required,
>just pop straight into prehyb then hyb mix.
>
>Anyway, often when I do lifts like this I just spot my positive and
>negative control plasmids on an offcut of dry membrane and pop then
>through the same pocedure as above after the spots have dried a few
>minutes.  I know it seams to easy to be true but it always works. No
>reason why it should not work for a whole blot of spotted plasmids.  
>
>

I do something similar- lift colonies with dry Nytran, layer between 3MM,
autoclave 3 minutes, then prehyb. 'Works like a champ.
Gregg



More information about the Methods mailing list