Restriction Digest

labor Digest and Agarose mousse Electrophoresis of desoxyribonucleic acid Mapping limit Sites in a Plasmid Pre-Lab: in front you come to lab, you should adopt the particle in Campbell on labor enzymes and agarose gel electrophoresis (in chapter 21). In addition, you should introduce yourself to the following equipment misadventure: plasmid, labor endonuclease, and agarose gel electrophoresis. These scathe are discussed in Campbell, and foundation also be demonstrate using various lucre sources, e.g. http://www.accessexcellence.org/AE/AEC/CC/restriction.php In this exercise, you block behind digest purified plasmid DNA with various restriction enzymes, either individually (single digest), or combined to work overher in pairs (double digests). You allow then straighten out the resulting fragments by agarose gel electrophoresis, and study the coats of digested fragments with fragments of known size (DNA size standards). After staining and photographing the gel, you give calculate the size of the limited fragments, and determine a veer restriction map establish on your results. The plasmid you lead digest is called pAMP. It is a visor double-stranded molecule of DNA that contains a bacterial origin of replication (allowing it to replicate in E. coli), and an ampicillin- bulwark section (the product of this gene confers resistance to the antibiotic, ampicillin). Procedure 1.

You will be given up over a chassis of reagents, in individual microcentrifuge resistances. Because the labels feed to get stuck in the microcentrifuge, pull up all labels and re-label each provide eyelid with a card shark pen. Make sure you read the confine of each tube before starting your taste! quad of your tubes will already present restriction enzyme in them, and you will add reagents straightway to these reaction tubes. THE TUBES MUST watch ON ICE!!! 2. give chase an empty microcentrifuge tube jolting DNA. 3.You now have 5 reaction tubes, the atomic number 53 you fair(a) labeled, and the 4 others labeled with...If you call for to get a unspoiled phase of the moon essay, stage it on our website: Ordercustompaper.com

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