HELP! Problems with RNA extraction from rabbit carotid

Geneboy Fiction at oz.net
Wed Jan 16 06:27:05 EST 2002


Hi Joyce,

I use TriZol to extract RNA from mouse brain and liver, with excellent
results. I homogenise the (snap-frozen) tissues with a blade homogeniser at
rt. As soon as I take a sample out of the -80C freezer, I pop it into the
TriZol and whizz. Works fine.

Maybe your problem is that you are going too close to the interphase when
you take up the upper (chloroform) fraction? I had several problems with
that, which I finally solved by not being too greedy, and leaving a
reasonable amount behind. Narrow tubes are good for this, as they minimise
loss.

I also use DEPC water throughout; I wash my tips and tubes in it too, and
then double autoclave them. I have a special set of pipettes for RNA work,
and so on. Typical RNA work precautions (although I have heard that they are
overrated).

HTH

Chris





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