lab Write-up - Jacob Zipperstein H. Bio Med January 28,...

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Jacob Zipperstein H. Bio Med January 28, 2009 Title : DNA Restriction Analysis ( Lambda DNA) Purpose : The purpose of this lab is to analyze Lambda DNA by using gel electrophoresis and different types of enzymes, which are known as restriction endonucleases. Procedures : The procedures that are undergone during this lab are actually quite simple. There are three particular endonucleases used in the lab: Eco R I, Hind III, and BstE II. The reason why we are using these endonucleases is because they can identify base sequences in the DNA. Once these sequences have been recognized, the enzyme will bond covalently to the Lambda DNA and will break the DNA bonds into specific regions. Once these “cuts” have been made, the Lambda DNA molecule will then easily break into many fragments. The first thing that needs to be done in order for all of this to happen is to load individual samples of Lambda DNA with a unique endonuclease. To do this, we are given three reaction tubes. A specific enzyme and a specific restriction buffer are assigned to each tube. Your job is too carefully load the correct amount of the buffer and the enzyme, using a micropipette, into one of the three tubes. Then, the same amount of Lambda DNA and the same amount of sterile dH 2 0 are added to all three tubes. Once each reaction tube has its specific enzyme and restriction buffer, then you take all three tubes over to be centrifuged. Then, you have to split up the tubes to be incubated. The tubes containing the EcoR I and the Hind III enzymes will be incubated at a temperature
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of 37 degrees Celsius, while the tube containing the BstE II enzyme needs to be incubated at a temperature of 60 degrees Celsius. The next step that needs to be done is to add dye into each reaction. Once you do
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lab Write-up - Jacob Zipperstein H. Bio Med January 28,...

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