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DNA extraction from paraffin blocks

song.65 at OSU.EDU song.65 at OSU.EDU
Fri Mar 12 16:32:12 EST 1999


My name is Nirav Shah, student at the Ohio State University.
I need help with DNA extraction from paraffin blocks.
I have tried this protocol but it didn't yield any products.
Anyone that can help me please e-mail me at      shah.198 at osu.edu

Here is the protocol I used:
1  Scrape off area of interest with a sterile (#11) scalpel blade and
transfer paraffin shavings
to sterile 0.5 ml tube.
2  Rinse shavings from blade with n-octane washes until there is 0.5 ml
in centrifuge tube.
3  Vortex for 20 minutes
4  Centrifuge for 5 minutes @ 6000 rpm
5  Pipette off n-octane using a pipetman with micro-capillary tip.
Usually, paraffin sticks to
sides of tube, avoid sucking it up.
6  Add 0.5 ml 100% ethanol to tube.
7  Vortex for 20 minutes
8  Centrifuge for 5 minutes @ 6000 rpm
9  Pipette off ethanol.  Avoid paraffin, which should be in a pellet at
the bottom.
10  Evaporate ethanol until pellet is dry several hours in a sterile
hood
11  Add 100ul digestion buffer (50mM Tris pH 8.5, 1mM EDTA, 0.5% Tween
20) and 2ul
Proteinase K stock solution (20 mg/ml in sterile H2O).
12  Place the sample tubes in a floating rack in a 55 C water bath.
Incubate for 48 hours.
13  Spin tubes briefly, then heat tubes at 95 C for 10 min to inactivate
the protease
14  Centrifuge at 6000 rpm for 1 min to pellet undigested material.

Thank you.
Again the e-mail address is shah.198 at osu.edu





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