Introductory Experiments in Recombinant DNA

Robert Campbell Tait

doi:10.21775/cimb.002.071

Abstract

Nine practical exercises demonstrate the basic principles in recombinant DNA. The exercises explain the principles that DNA equals genes and that changes in DNA cause changes in genetic properties. The aim is to provide a teaching resource that can be used to illustrate the theory and applications of molecular biology to highschool students, undergraduate students, medics, dentists, doctors, nurses, life scientists, and anyone learning the basics of DNA technology.

Highlights

Nine practical exercises demonstrate the basic principles in recombinant DNA
The aim is to provide a teaching resource that can be used to illustrate the theory and applications of molecular biology to highschool students, undergraduate students, medics, dentists, doctors, nurses, life scientists, and anyone learning the basics of DNA technology
The exercises contained in this article have been chosen to demonstrate the basic principles in recombinant DNA: digestion of DNA with a restriction endonuclease, gel electrophoresis of DNA samples, insertion of DNA into cells can change their growth characteristics, and DNA can be rearranged to cause changes in genetic properties

Summary

Introduction

The exercises contained in this article have been chosen to demonstrate the basic principles in recombinant DNA: digestion of DNA with a restriction endonuclease, gel electrophoresis of DNA samples, insertion of DNA into cells can change their growth characteristics, and DNA can be rearranged to cause changes in genetic properties. Nucleic acid samples are often subjected to gel electrophoresis to characterize the size and number of different fragments in the sample In this exercise, DNA samples that have been digested with restriction enzymes and mixed with a tracking dye will be subjected to agarose gel electrophoresis. The result is that molecules separate in the matrix according to their relative size and shape This exercise will use gel electrophoresis to examine the fragments present in several DNA samples. Several DNA samples from genomes of increasing size will be digested with EcoRI in this exercise If both the enzyme digestion and the gel analysis are to be performed in the same day, as soon as the digestion reactions have been set up and are incubating, prepare agarose minigels for analysis of the digested DNA samples. They are the undigested controls and will be used to compare to samples following digestion

To the remaining 40 μl of each sample add

Label four of the capped sterile glass or plastic tubes as below

Background

Load the samples in the following order and amount