isolation of DNA from mouse tails

Ulrich Tillmann mcarb at pop.erols.com
Wed Jul 24 09:37:24 EST 1996


Dear John: Here is a protocol I have used successfully on  thousands of 
tails (PCR, Southern etc.).


Lysis solution: 200 mM NaCL, 20 mM EDTA, 40 mM Tris-HCL ph8, .5% SDS,.5% 
Mercapto OH(fresh!!)

Saturated NaCL (approximately 6 M): add solid NaCL to 5 M NaCl stock 
solution


1) make little incisions on tails and put in lysis solution with 200 ug/ml
 
Prot. K.

2) Incubate o/n or shorter at 50 C.

3) After digestion add 0.5 vol saturated NaCl, mix carefully and stand on 
ice at least 10 min.

4) Spin in tabletop centrifuge

5) transfer supernatant with wide bore plastic pipettes into clean tubes 
and precipitate DNA with 2 vol of 100% EtOH.

6) Collect pellet, wash in 70% EtOH and dissolve in TE buffer pH 8.




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