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>
