1)Aliquot5X106totalcellsin5-8ml2%media.2)Add30µl3H-thymidine.3)Incubateforapproximately16hoursundernormalgrowthconditions.4)Spindowncellsandwash2-3timeswithPBS(orserumfreemedia)toremoveanyunincorporatedlabel.5)ResUSPendcellsat5X105/mlinserumfreemedia.6)Aliquotapproximately400µlofcellsuspensionintoeachwellofa24wellplate.7)Treatcells(ALWAYSHAVETIME-MATCHEDCONTROLS!!).8)Pelletcellsgentlyandcountsupernatant.-->supernatant=A9)Resuspendpelletinlysisbuffer.10)Microfugecellsuspensionfor15minutesatmaximumspeed.11)Collectpelletandsupernatantandcountboth.-->supernatant=Bpellet=C12)Todeterminethepercentfragmentationusethefollowingequation:-->(A+B)/(A+B+C)
Lysisbuffer