requested protocol for tail snip DNA

Darin Obrien IHG darin at gene.med.umn.edu
Sat Jun 17 17:17:56 EST 1995


Hi folks,

	I have had a number of requests for this protocol since 
mentioning it a couple weeks ago so I thought I would go ahead and post 
it.  So here it is.  If you have any questions, feel free to contact me.

best of luck,

darin obrien
darin at gene.med.umn.edu


DNA Isolation from Mouse Tail Snips


 1.  Add 10mm tail snip to 1.5ml microcentrifuge tube.

 2.  Add 0.6ml TES.

 3.  Use pestle to grind tissue (approximately 10 strokes).

        NOTE:  these little blue pestles can be purchased from VWR.  I can
 find the catalog number if you need.

4.  Incubate at 65oC for 15-30 minutes.

 5.  Add 17ul Proteinase K solution (20mg/ml).  Mix the sample by
 inverting the tube 25 times.

 6.  Incubate at 55oC for 2 hour with shaking.

 7.  Add 6ul RNaseA solution (10mg/ml).  Invert the tube 25 times to mix
 the sample.

 8.  Incubate at 37oC for 30 minutes.

 9.  Add 0.2ml 5M NH4OAC and vortex vigorously 20 seconds.

 10.  Centrifuge at maximum speed in microcentrifuge for 10 minutes to
 pellet proteins.

 11.  Pour the supernatant into a fresh 1.5ml microcentrifuge tube and add
0.6ml isopropanol.

 12.  Invert the tube 25 times to precipitate the DNA.

 13.  Centrifuge 15 seconds in the microcentrifuge at maximum speed to
 pellet the DNA.

 14.  Wash the pellet one time with 200ul 70% ethanol (rock the tube until
 the pellet is no longer stuck to the bottom of the tube).

 15.  Pellet the DNA by centrifugation for 15 seconds.

 16.  Discard supernatant and allow to air dry for 15 minutes.

 17.  Add 500ul TE and rock overnight to resuspend.


 NOTE:  yields vary considerably but 150mg yield is average.
 Concentration will be ~ 0.3mg/ml.
 TES = 10mM Tris-HCl, pH 8.0, 25mM EDTA, 0.5% SDS.
 5M NH4OAC = 19.27g in total volume of 50ml.



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