Protocol for PCR able DNA extraction from Wax-embedded tissue and Formalin fixed tissue.

1 mbad at magnum.barc.ernet.in
Thu Jan 15 23:16:45 EST 1998


Fellow workers,
	I am working on Microsatellite instability in Thyroid Cancer tissues.
I need to protocols to extract PCR-able DNA from Wax Embedded Tissues and 
Formalin fixed tissues.Can Anyone direct me any such references and 
protocols both in print and on the Net if available.
	If anyone is working on similar lines I would be grateful if 
could send me some tips on better amplification from such archival tissues.
	Presently The protocol I use on wax section -- dewaxing with 
xylene-3x,Digestion with 50mM tris(pH8.5)and 1mM EDTA and 0.5%Tween 20 
with 200ug/ml Protienase K at 55 degrees C overnight.
(Reference:PCR protocols- A guideto methods and applications edited by 
Michael A. Innis etc.. publisher- Academic Press,Inc)
	
	I used this supernatant for PCR but it doesn't amplify - a faint 
smear along the length of gel is visible.
	Can anyone help.
	Suresh Shettigar,
	Scientific Officer,
	1-108-H,MB&AD,
	Bhabha Atomic Research Centre,
	Trombay,Mumbai-85.
	INDIA.
email:<mbad at magnum.barct1.ernet.in>



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