Wedge Sequencing gels

Andy Phillips andy.phillips at bbsrc.ac.uk
Wed Aug 14 02:56:48 EST 1996


Dr. Duncan Clark wrote:
> 
> Hi Folks,
> 
> We are using a Stratagene Base Ac jnr sequencing set up with 41 x22cm
> plates. If we want to make wedge shaped gels any guide on both waht
> thickness of spacer (two or three thicknesses?) we should use at the
> bottom and when ie how far up the gel do you jump to the normal spacer.
> We are currently using 0.4mm spacers but can go down to 0.2mm. On this
> length of gel are wedge gels of use anyway?
> Duncan, 

Don't bother with wedge gels. A far simpler method for slowing down the bottom 
half of the gel is to add 1/6 vol of 2M NaAc to the bottom revervoir after 
you've loaded the samples. If you're doing a long run, add it about halfway 
through. This increases the conductivity of the bottom of the gel and so slows 
down the DNA fragments. Some people use 1/3 vol of 2M, but I've never tried 
this.

Andy
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