Age and Page

AGEagaiose - uncharged linear polymer made up of repeatingunit of agaroblose disacchaide made up of D-galaclose and 3, 6 - arhydeo - L- galactopyrancleThe gel is prepared in buffer and spread over microscopicplateA small sample is applied by cutting into thegel with a shoup edge by using combining techniques.The electrophoretic rurs usually takes 2 90 mins.The migration of nucleic acids in agaiose gels aalso affected by the choice of running buffer and theapplied voltage.widely used for separating NA.Simple + nontoxic offer separation rangesize of gel pores can be controlled by simplyadjusting the agarose Concertisation to prepare gels appropriatefor the separation of a wide range of different size Nuclic acid molecule40.5I - 30 kb 1.2400bp - 7kb0.7 800p 12kb 1.5 200bp - 3 kb/500bp 10kp 2 so bp - 2k b./ PAGEprepared by Co -polymerizing the acrylanide and methylere bisanylamicle to Covalently crosslink the monomersThe reaction is iniated by a free radical generaling systemAPS and TEMGD.Tougher than agarose gel.Thumostable, transport , relatively chemically inusCuls are uncharged and can be prepared in variety ofpore sitesAgaiosePAGEHorizontalverticalseparate large molsmall mols.DNA separationPro separationStairing car be donestairing after pouringthebefore pouring the gelgelCapillary electro phoresis :-A typical capillary electrophoresis system consists of a high vottagepower slipply, a sample in troduction system ,a capillarytube , a detector and an output device.Some instruments include a temp control devicetoensure reproducible results. This is becoz the separationof samples depends on the electropheratic mobility andthe visiosity of the solutions Yes as the column temp1-1 Capillary electropheresislight sourceaM=16 = GITHYcomputerBuffeadjerBathodetillAnode (+)(-)with capillary all analytes travel the same distancebut the migration time (t m) for that distanceis measured.Relate time to identityRelate peak area or height to amountEach dide of high voltage power Supply is connectedto an electrode. There electrodes hilp to induce anelectric field to initiate the migration of thesamplefrom the anode to the cathode through the capillary tube.The capulary is made of fused Solica ay ll sometimescoated with polyimideEach side of capillary like is dipped in a vial containingthe electrode and an electrolytic d dn or aqurous buffer.There is usually a small window near the cathodic end ofthe capillary which allows UV-VIS light to pass throughthe analyte and measure the absorbance.CE tubes have only small amount of running buffer(0.5mL)coupled with high efficacy only a small sample sizeCan peinjected. Hydrodynamic injection. - press wike the sample condainingContainer after connecting it to the capillary true thenrelease and replace the capillary tube in the running bufferElectroknitic injection :- electrode and capillary all incontact with the sample - the applied field cause theSample to enter the tube -CG had showed solved some common problems into aditional electrophoreisThe thin dimensions of the capularies greatly redthe & you & to volume ratio which deminatedoverheating by high voltages.CE uses tubes 20-100 mm in d as 20-100 cm in l.using narrow bore twices LG removes the Joule heatingeffect which 2 band broadering giving fasterSeparations than gel.often CE is used with no gel (no eddy diffusion)LongiEudiral diffusionmain Source of bandbroaderingMoving boundry Electrophoresis 3-developed by A. Tosclius of SWEDEN in 1934Electrophoresis is cavid in Solution without a supportingmediaSample disolved in buffer and molecules move to thatrespective Courtes change electiode.MBE is carried out in a ushape like withplatinum electiocles attached to the end of both carms.