Methodology and Equipment for General Molecular Techniques

The following sections are included:

• Introduction

  • DNA Extraction

    • CTAB Extraction [modified from Draper and Scott (1988)]

    • Potassium Acetate/SDS Precipitation Method [modified from Dellaporta et al. (1983)]

    • RNase A Digestion

    • DNA Concentration/Purity Estimation

  • Electrophoresis

    • Agarose Gel Electrophoresis of DNA

    • Denaturing PAGE

  • Purification of PCR Fragments Using a GENECLEAN III Kit (Bio 101 Inc.)

  • DNA Modification

    • End Modification of PCR Amplified DNA

    • Radiolabelling of Oligonucleotides

  • Cloning

    • Ligation and Transformation of E. coli

    • Screening for the Presence of Insert in Plasmid by Rapid Alkaline Lysis (Barnes, 1977)

  • Cycle Sequencing of Plasmid DNA

  • Similarity Index (S.I.) Analysis

• References