IsolationofDNAfromMouseTailBiopsies 1.Obtaintailbiopsiesfrom2to3weekoldmice:Holdmousefirmlyatbaseoftailwithonehand,withtheothercutoff0.5to1.0cmofthetailtipwithascalpelorsingleedgerazorblade. 2.Add600microlitersofTNESand35microlitersProteinaseK(10mg/ml). 3.Incubateovernight(8-24hr)at55degreesC. 4.Add166.7microliters6MNaCl.Shakevigorouslyfor15sec. 5.Microfuge(12,000-14,000xg)for5minatroomtemp. 6.Removesupernatanttonewtubeandadd1volumecold95%ethanol(EtOH). 7.SpoolprecipitatedDNAwithclosedendcapillarytube. 8.RinseDNApelletwith70%EtOH(dippinginatubefilledwith70%EtOHworkswell)andallowtoairdry5-10min. 9.ResUSPendin100-500microlitersofTE(10mMTris,pH8.0,1mMEDTA).Volumedependsempiricallyonpelletsize. 10.Heatat65degreesCfor10minutestoaiddissolutionofDNA. 11.QuantitateDNAandstoreat4degreesCuntilneeded. "6M"NaClSaturatedsaltsolutionstoredat37°C MillerSA.DykesDD.PoleskyHF.1988.AsimplesaltingoutprocedureforextractingDNAfromhumannucleatedcells.NucleicAcidsResearch.16(3):1215.TNES10mMTris,pH7.5400mMNaCl100mMEDTA0.6%SDS
ThereferenceforthetailDNApreparationis:
