Lab 6 Restriction mapping

Lab 6 Restriction mapping - Q1(2pts Why is it necessary to...

Info iconThis preview shows pages 1–5. Sign up to view the full content.

View Full Document Right Arrow Icon

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: Q1 (2pts): Why is it necessary to cut a plasmid with a restriction enzyme before its size can be determined by agarose gel electrophoresis? Q2 (3pts): What information about a plasmid do we need in order to confirm its identity accurately by restriction mapping? Why? Lab #6 Plasmid Identity by Restriction Mapping Objectives: • to run agarose gel electrophoresis of restriction enzyme digested pGEM and pGEM- luc • to analysis restriction digested fragments pellet cells pGEM pGEM- luc restriction digest Plasmid isolation Approaches to Confirm Plasmid Identity 1. Restriction enzyme digestion followed by gel electrophoresis of digested fragments 2. DNA sequencing Last Week: 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. Predicting restriction digest patterns from a...
View Full Document

This note was uploaded on 09/12/2009 for the course BISC 320L taught by Professor Baker,aparicio during the Fall '07 term at USC.

Page1 / 10

Lab 6 Restriction mapping - Q1(2pts Why is it necessary to...

This preview shows document pages 1 - 5. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online