An experiment aimed at performing electrophoresis using restriction enzymes and lambda dna

Why do you heat the lambda dna fragments prior to electrophoresis

Add enough methylene blue DNA stain to cover the gel and place cover on it. Restriction enzymes play a very important role in the construction of recombinant DNA molecules, as is done in gene cloning experiments. Steady pipette over well using 2 hands. Note Gels may be discarded in regular trash receptacle. Let cool until solidified approximately 15 minutes. The gels can then be placed on gel support film, which binds the gel and dehydrates it, if your instructor so chooses. Make certain that sample wells left by comb are completely submerged by buffer.

If it is present, put the sample back and begin again. Objectives Understand what a DNA restriction enzyme is and how it works. The fragments move through the gel at a rate that is determined by their size and shape, with the smallest moving the fastest. Learn to separate DNA on an agarose gel using electrophoresis.

How long will a fragment of lambda dna produced by a sau3ai digest be

Electrophoresis The electrophoresis chamber top is placed on the chamber, the electrodes connected to power supply--anode to anode red-red and cathode to cathode black-black. Background Reading Since viruses have a relatively simple genome, scientists have studied their DNA and used this information to test theories and develop concepts that apply to the genetics of living organisms. Slide gel easily into staining tray labeled with your group name. Otherwise, allow the solution to cool and solidify. It will initially appear as a blue band, eventually resolving into two bands of different colors. Dip pipette tip through buffer, positioned over the well, and slowly expel the mixture do not punch thru bottom of gel. The gels will be washed a couple of times in distilled water standing for min , and will refrigerate until next lab period when we will look at them. After the sample is ran, the unknown fragments can be compared with the ladder fragments to determine the approximate size of the unknown DNA bands by how they match up to the known bands of the ladder. Carefully remove the casting tray and keep it horizontal until you are ready to put into the plastic container. The slower moving aqua band is xylene cyanol, nearly equivalent to a base pair fragment. Do NOT move tray while agarose is solidifying. How can you account for differences in band separation and intensity between your gel and the ideal gel? Use the stain close to a sink and clean up spills immediately. Add enough methylene blue DNA stain to cover the gel and place cover on it. You now have a sample inside of your pipette tip.

Add enough methylene blue DNA stain to cover the gel and place cover on it. How can you account for differences in band separation and intensity between your gel and the ideal gel?

restriction enzyme digestion lab report answers

The correct restriction maps may be viewed on-screen. Place the plastic comb in the slots close to the end of the tray.

Restriction digestion of lambda dna

You now have a sample inside of your pipette tip. Gel will solidify within 20 minute. Place the plastic comb in the slots close to the end of the tray. Slowly depress plunger to first stop and then depress to second stop to blow out last bit of fluid in tip. A single container of methylene blue dye should be all that is needed since it may be reused several times and disposed of down the sink. Use the stain close to a sink and clean up spills immediately. You will need to remember the order of your tubes since there is NO way that the gel can be marked. Dip pipette tip through buffer, positioned over the well, and slowly expel the mixture do not punch thru bottom of gel. The gel is now ready to load with DNA.

Electrophoresis The electrophoresis chamber top is placed on the chamber, the electrodes connected to power supply--anode to anode red-red and cathode to cathode black-black.

They lose activity unless kept frozen; exposure to warm temperatures for even a short time will result in loss of activity. Use pipette to load contents of each reaction tube into a separate well in gel total of 4 wells.

This step eliminates any air in tip. Fill box with TBE buffer, to level that just covers entire surface of gel by about 2 mm. Use a clean pipette tip for each different tube.

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DNA Restriction and Electrophoresis