BLUE HAZE

Dr Mick Jones mjones at rpms.ac.uk
Mon Apr 20 11:32:56 EST 1998


Hi out there

I apologise for asking about BLUE HAZE problems, but until recently we have
never had them (and we have been using a 373A sequencer for about 5 years). 
I tended to look at the emails about blue haze and forget them.

Last month we changed the filter wheel on our 373A Stretch XL instrument
(upgraded last November) to enable us to take advantage of the big-dye
terminator (BDT) kits.

On our third run with the BDTs we experienced a blue haze over the gel, and
we still have the problem.  Although we can raed the trace underneath
reasonably well.

We just ethanol ppt our reactions prior to dissolving in loading buffer
(stored at -20C), heat in an oven for about 5 minutes at 80 - 90C, and then
load immediately.  The plates are just washed in hot tap water after the
run, and then washed in deionised Millipored water and dried with a car
rubber windscreen wiper before pouring the gel.

Is the haze due to something in the water (revealed now because of the
changed filter wheel) or due to degrdation of the formamide loading buffer
or is there something in the BDT kits?  It does seem strange that we never
had the problem, and when we changed the filter wheel and started with the
new kits we get the blue haze.

Many many many thanks for your replies.

Mick Jones

PS please email as well as post.  Thanks.
 
Dr Mick Jones, Lecturer in Molecular Virology
Department of Infectious Diseases and Microbiology, Imperial College School
of Medicine
Hammersmith Hospital, Du Cane Road, London, W12 0NN
Tel:  0181-383-3328;  Fax:  0181-383-2299
email:  mjones at rpms.ac.uk
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