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Mouse Tail Genomic DNA Isolation Protocol(1)

n carefully transfer resultant aqueous phase to a fresh microcentrifuge tube.
  • Fill sample-containing tube with 100% Isopropanol and mix thoroughly by repeated inversion. Do not vortex. A visible DNA precipitate should form. Proceed immediately to step 11.
  • Recover DNA precipitate by touching it to a heat-sealed glass micropipette tip or by lifting the DNA with a yellow pipette tip and partial suction from a pipettor. Transfer the DNA to a 1.5 ml microcentrifuge tube containing 70% Ethanol. If DNA is not stringy, pellet by a brief, low speed centrifugation.
  • Wash DNA with the 70% Ethanol, then wash twice with 95% Ethanol.
  • Allow DNA to partially dry and then either transfer the DNA to a microcentrifuge tube containing 400 l TE (10 mM Tris-Cl, 1 mM EDTA, pH 8.0) or add 400 l TE to the DNA in the tube. Do not vortex or re-pipet to resuspend DNA.
  • Resolubilize the DNA overnight (e.g. by rotation at 3060 rpm). Resolubilization may be facilitated by heating the sample at 50C.
    • References
    1. Herrmann, B.G. and Frischauf, A.M. (1987) Isolation of genomic DNA. Methods Enzymol. 152:180-183.
    2. Laws, G.M. and S.P. Adams. (1996) Measurement of 8-OHdG in DNA by HPLC/ECD: The importance of DNA purity. BioTechniques 20:36-38.
    3. Murphy, N.R. and Hellwig, R. J. (1996) Improved nucleic acid organic extraction through use of a unique gel barrier matrix. BioTechniques 21:934-939.
    4. Fil Randazzo, personal communication.

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