Purpose: Timerequired: Day1-3:10minutesDay4:6-8hoursDay5:2hoursDay6:1hour SpecialEquipment: SpecialReagents: Procedure: Days1-3 Day4 Days5-6 Solutions: 3%Sarkosyl 0.2MEDTA References: Carle,G.F.,andM.V.Olson.(1984)"SeparationofchromosomalDNAmoleculesfromyeastbyorthogonal-field-alterationgelelectrophoresis."NucleicAcidsRes.12:5647-5664. Burke,D.T.,Carle,G.F.,andM.V.Olson."CloningoflargesegmentsofexogenousDNAintoyeastbymeansofartificialchromosomevectors."(1987)Science236:806-812.Toisolateintact,highmolecularweightDNAfromyeastcellsforsubcloningandrarecuttingrestrictionenzymeanalysis.Onecanexpectayieldof100-200礸ofDNAperprep.
6daystotal
Filtersterilizeandstoreatroomtemperature.SCE Finalconcentration 2MSorbitol 50ml 1.0M 1MSodiumcitrate 10ml 0.1M 0.25MEDTA,pH7.0 24ml 60mM sterileddH2O 16ml ------- 100ml 0.5MTris-HCl,pH9.0