Manual

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SECTION 4
Running Conditions
4.1 Recommended Power
Therecommendedpowerconditionsforoptimalresolutionare50to150V,constant,
and25to80mA.Theusualruntimewillvaryforthevoltagechosenbutshould
rangefrom15to60minutes,monitoringnucleicacidmigrationbyprogressofmarker
dyes.Constantpowerisnotanecessity,butitproducesuniformheatthroughoutthe
run.Besurethepolarityiscorrecti.e.thattheDNAisloadednearthecathode(black
electrode)toruntowardtheanode(redterminal).
Agarosegelsmaybestoredforseveraldaysat4°Cwrappedinplasticwrap.Seakem
Agarose(FMC)isused(normally)forpreparativeandanalyticalgels.Othertypesof
agarosecanbeusedforspecialpurposes.
4.2 Recommended Buffers
Type  ConcentratedStock/liter  FinalConcentration
TAE  50X-242gmTrisbase  1X-0.04MTris-acetate
(Tris-acetate) 57.1mlglacialaceticacid  0.001MEDTA
  100ml0.5MEDTA(pH8.0)
TBE  5X-54gmTrisbase  0.5X-0.045MTris-borate
(Tris-borate) 27.5gmboricacid  0.001MEDTA
  20ml0.5MEDTA(pH8.0)
10X-LoadingBuffer(DNA)
0.25%Bromophenolblue
0.25%Xylenecyanol
20%FicollType400
0.1MEDTA,pH8.0
4.3 References
1)Hames,B.D.,Rickwoood,D.(ed.)(1990).GelElectrophoresisofNucleicAcids.APracticalAp-
proach.2ndedn.IRLPress,Oxford.Ch2.
2)Sambrook,J.,Fritsch,E.F.,Maniatis,T.(1989).MolecularCloning.ALaboratoryManual.2nd
edn.ColdSpringHarborLaboratoryPress,ColdSpringHarbor,NewYork.Ch6.
3)Ausubel,F.M.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.,Struhl,K.(ed)
(1993).CurrentProtocolsinMolecularBiology.Vol.1,GreenePublishingAssociates,Inc.and
JohnWiley&Sons,Inc.,Ch.2.