University:
Swansea University
Course:
PM134 | Microbiology
Academic year:

2023
Views:

52

Pages:

2
Author:

Sanjeev K

EXPERIMENT NO:3DATE : 13.10.20225AGAROSE GEL ELECTROPHORESISAIMTo separate DNA samples on the basis of their sizeusing agarose gel cleutrophoresis techniquePRINCIPLErding to their rize. DNA negatively In the presence of anAgarose gel elutrophoresis is separates DNA charged fragments due auo. toelectric the presence field, end of the phosphate - vely the field charged Agarose DNA migrates is a linear which polysacchan is extractgroups.towardsthe made positive up of repeating of units of agarobiose,from certain The separation seawads of DNA smaller fragments molecules depends moves upon faster theconcentration than ones of against Unknown The fragments are to compared determine withthe larger fragments, whose size is known bandsthe ladetop approximate Size of the unknown DNAAgarose, TBE buffer, gelcasting Bromide, tray, comb, DNA power ladder, pack, micropi- SampleMATERIALS REQUIREDDNA, comical loading flask, dye, microwave Ethidium oven, tank Gel documentationsystem, Pette, cellotape, electrophoretic300 PROCEDURE mg of 1% Agarose is prepared in 30ml TBE bufferin a comical flaskThe comical flask is then kept in the microwave ovenAs soon as the comical flask is taken from the oven,for a few minutes to dissolve the agaroseEtBr is added.Meanwhile, the gel casting tray is kept ready by is wrappingits edges using cellotape and then the comb placedwhen the agarose gel reaches a bearable temperatureIt is then kept for solidificationIt is carefully poured in to the gel casting tray. 6As soon as the solidification is completed, the tray isPland inside the electrophoretic tank containing thesame TBE buffer after removing the cellotape.The camb is removed and 2011 of ladder DNA iscarefully loaded using a micropipette in the first the well.The provided unknown DNA sample mixed withloading dye is then loaded in the other that wells It (2011) is closerto The carling tray is kept in such a wayThe red lead of the tank is plugged in the red jack is andnegative electrode.black lead in to the black jack, which in turnconnected to the power pack.with 50 V supply for the first five minutes, men gradu-The lid of the tank is placed and power park is switchedorally it is increased to 100 and 150Vwhen the DNA migration reaches about 3/4th of thegel matrix, the power supply is suitched off It isunder the get documentation observed system, where differentthen carefully lifted from the tank and examinedRESULTSlength," DNA fragments isThe given unknown sample is separated and observedgot