Lab _6 Restriction mapping friday

Lab _6 Restriction mapping friday - Lab #6 Plasmid Identity...

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Lab #6 Plasmid Identity by Restriction Mapping Section 1332 7 Friday 1:00pm-4:00pm ZHS 359 Zheng Zhang Office hours: T/Th 11:00am-12:00 zhengz@usc.edu
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Objectives: to run agarose gel electrophoresis of restriction enzyme digested pGEM and pGEM- luc to analysis restriction digested fragments Pour 0.8% agarose gel at the beginning of lab. http://www.gbiosciences.com/EducationalProducts/Plasmid-Isolation-(Alkaline-Lysis).aspx
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pellet cells pGEM pGEM- luc restriction digest Plasmid isolation Approaches to Confirm Plasmid Identity 1. Restriction enzyme digestion follow by gel electrophoresis of digested fragments 2. DNA sequencing to check whether there are mutations in the cloned gene Last Week:
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3 major forms of plasmid DNA C. Supercoiled: compact shape, no excessive damage (fastest) A. Relaxed or Nicked Circular: nicks (on one strand of DNA) in the supercoiled plasmid (slowest) B. Linear : direct opposite strand nicks or cuts by restriction enzyme C. Highly supercoiled: has more superhelical turns The size of DNA is determined by looking at its linear form and compare it to a DNA marker.
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This note was uploaded on 12/01/2009 for the course BISC 13019 taught by Professor Aparicio during the Spring '09 term at USC.

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Lab _6 Restriction mapping friday - Lab #6 Plasmid Identity...

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