Gelelectrophoresis separates DNA fragmentsaccording to sizeElectrophoresis: The movement of charged molecules in anelectric field.Preparing an agarose gel for electrophoresis.Amachacombtoackoaracrylic2.Pourhealodnoitenaarosintoplate3.Removecoeribfroengel:plate with clamosAllowtocodiardhatdonshallcwweisareloft in DelRemovegetfron pliate.9-16Fig.9.4a
9-16 Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display Gel electrophoresis separates DNA fragments Gel electrophoresis separates DNA fragments according to size according to size Preparing an agarose gel for electrophoresis Fig. 9.4 a Electrophoresis: The movement of charged molecules in an electric field
Loading DNA fragments onto an agarose gel and performingelectrophoresis.AmicropipetfeisusedtoloagDnAsarnplesintoeachwedS.EoctrodowiresarepitcedalorgeachendofteoeEachsarnplecontainsatkvedyetomakeiteasiertoseeandareatachedtoapowersuooly.TheCurteritissvilchedon,andCnAmolecuesineachsamolmigrate toward theend ottebox (along the tathisCepictedwithorangearrows).Eectrophoresilsconeinuesfot1-20hcurs9-17Fig.9.4a
9-17 Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display Loading DNA fragments onto an Loading DNA fragments onto an agarose agarose gel and performing gel and performing electrophoresis electrophoresis Fig. 9.4 a
Visualizing the DNA fragments on the agarose gel6,Removegetfrom get ticx,tncubale with ethidiambramide.then.washto-removo ewcessdye.BrCHsethidium bromide溴化乙锭9-18Fig.9.4a
9-18 Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display Visualizing the DNA fragments on the Visualizing the DNA fragments on the agarose agarose gel Fig. 9.4 a 溴化乙锭
Polyacrylamide gels separate very small fragments ofDNA and agarose gels separate larger fragments404kb30923242-1806.6-147~4.41232.32.0-90.9-19Fig.9.4b
9-19 Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display Polyacrylamide Polyacrylamide gels separate very small fragments of gels separate very small fragments of DNA and DNA and agarose agarose gels separate larger fragments gels separate larger fragments Fig. 9.4 b
Find landmarksinDNA byrestriction enzymesRestriction maps can beinferred fromDNA fragments cutwithtwo enzymes providing a roadmap to DNA fragments and virusgenomes.EcoRI15.4/0Clal3.KpnI KpnISall600450200400EcoRV11.951650Hindill11.54.011.3HindiIllEcoRVpss2 (15.4 kb)4.4.Bglll4.4510.76EcoRVEcoRV5.59.79.256.25XholClal9.06.857.66BgillXholBgllEcoRVEcoRV9-20
9-20 Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display Find landmarks in DNA by restriction enzymes Find landmarks in DNA by restriction enzymes Restriction maps Restriction maps can be inferred from DNA fragments cut with can be inferred from DNA fragments cut with two enzymes providing a roadmap to DNA fragments and virus two enzymes providing a roadmap to DNA fragments and virus genomes. genomes