Agarose Gel Electrophoresis

this video will take you through the process for loading and running DNA samples on an agarose gel you will be using a mini-sub cell to perform agarose gel electrophoresis the sub cell has an electrode wire running across the bottom of each end this provides from electric current to pass which separates the DNA fragments in the samples first align the gels so that the wells are closest to the negative or black electrode DNA is negatively charged and will move from the negative electrode through the gel toward the positive or red electrode place the agarose gel into the gel chamber add electrophoresis running buffer to the reservoirs at each end of the gel chamber and keep adding buffer into the wells in the gel are covered by at least two millimeters of buffer you place your samples to be loaded in the correct order according to the lanes they are assigned to be running check your lab protocol for this information to obtain a DNA sample slowly depress the plunger on your adjustable micropipette to the first or soft stop holding the plunger down insert the tip of the pipette into the micro tube with a DNA sample place the tip close to the bottom of the sample then slowly release the plunger button when loading the samples keep the pipette tip perpendicular to the row of wells this will reduce the risk of accidentally puncturing the wells with a tip lower the tip of the pipette until it breaks the surface of the buffer and is located just above or just inside the well slowly apply pressure to the plunger button observe as a sample fills the well make certain you stop at the first stop on the pipette pause then slowly remove the pipette while keeping your thumb down on the plunger once all the samples have been loaded avoid any bumping or movement of the gel chamber this might result in the sample spilling into adjacent wells place the lid on the gel chamber with the terminals correctly positioned to the matching electrodes on the gel chamber black to black and red to red connect the electrodes to the power supply again making certainly electrodes match the terminals on the power supply red to red and black to black switch the power supply on then set the correct constant voltage for running the samples see your lab protocol for this information if a timer is available on your power supply set the clock for the proper time for your run press the start button to begin the flow of current that will separate the DNA fragments at this point you should be able to see bubbles coming from the wires at each end of the gel box notice that the positive or red electrode has fewer bubbles than the negative or black electrode in a few minutes you should see the samples begin to migrate from the wells into the gel notice as your DNA samples run the diaphragm moves from the negative electrode towards the positive electrode you

Channel: Bio-Rad Laboratories

Views: 1,447,170

Keywords: bio-rad, life science, education, biotechnology, biology, molecular biology, technique, method, student, teacher, school, classroom, laboratory, laboratory skills course, textbook, lab, activity, demo, demonstration, instuctional, instruction, agarose, gel, electrophoresis, dye, dye electrophoresis, loading, load, pipette, bromophenol, blue, separation, run, running, kit, idea kit, stem, dna, dna structure, ngss

Id: vq759wKCCUQ

Length: 4min 7sec (247 seconds)

Published: Fri Oct 12 2012