RNA isolation from PANCREAS

POPOVA ANDREI.POPOV at afrc.ac.uk
Thu Dec 1 07:17:58 EST 1994


In Article <D0371C.8wI at liverpool.ac.uk>, ferret at liverpool.ac.uk (Mr J.S.
Erikson) wrote:
>
>Hi,
>        Am I having major problems with this one.
>Ive been isolating RNA from tissues and from leukocytes using several diff
>erent methods; Trizol(Gtc complex [GIBCO], Lithium chloride/urea; and alkaline
>lysis methods.  Recently Ive tried to isolate total RNA, with the hope of
>performing Northern hybridisation from PANCREAS.
>
>Q.      The surgeon who is excising the pancreas has been snap freezing them
>in liquid nitrogen, will this fragment mRNAs of approx 1-2kb?
>
>Q.      Would it help to excise the pancreas and immediatley homogenise in
>the isolation agent?
>
>  Unfortunately Im losing the RNA through degredation.  Ive had an internal
--------------------------------------------------------------------------------
>control in each extraction (Human P.B.L.s) to check my technique is ok.
>
>        Any good ideas out there?  e-mail: ferret at liv.ac.uk
>
>                        Thanks people  John Erikson  

HI JOHN
I DID NOT UNDERSTAND WHAT YOU MEAN BY 'INTERNAL CONTROL'
DOES IT MEAN YOU ADD P.B.L. INTO PACREATIC TISSUE?

I WOULD SUGGEST TO USE FRESH TISSUE AND USE 6.5M GIT INSTEAD OF
4M. THIS CHANGE HAS BEEN DOCUMENTED (NAR, ?????) FOR USE
IN CHOMSZYNSKY AND Co. PROTOCOL AND GIVES BETTER RNA RECOVERY
FROM DIFFICULT SAMPLES
I USE 6.5M GIT ROUTINELY FOR ALL RNA EXTRACTIONS

BEST WISHES
ANDREI POPOV




More information about the Methods mailing list