Report on Recombinant DNA Techniques (Micromon) - Heart Research Institute
S Nakhla
Senior Research Assistant
Clinical Research
The Heart Research Institute
I was very fortunate to attend an intensive Recombinant DNA Techniques Course at the Department of Microbiology, Monash University Clayton, Victoria, from Sunday November 18 to Friday November 23, 2007, as the recipient of a $1600 ASI study grant.
The course provided comprehensive and fundamental training in the essential skills of “recombinant DNA technologyâ€.
Recombinant DNA technology is a technique in biotechnology which involves the artificial re-arrangement of DNA by isolating segments of DNA from one organism, which can then be incorporated into the genetic makeup of another organism.
The aim of this process is to mass produce the protein encoded by the inserted gene along with substances coded for by the native genetic material of the recipient.
The techniques involved have become well established and are universally applied to solve biological problems particularly in the diagnosis of genetic disorders and determination of potential causes of many diseases. These techniques have transformed the field of research in the biological and medical sciences.
The course format was ten hours of theory via lectures and over thirty hours of practical training which involved experimental laboratory work and tutorials.
Lectures included a basic introduction to Microbiology, DNA and Molecular Biology.
Basic Cloning Requirements, Gene Cloning Techniques and Hybridisation – Northern, Southern and Western blots.
The theoretical basis and applications of construction of Genomic Libraries, Restriction Fragment Mapping, Genetic Mapping, DNA Sequencing Design, the use of Oligonucleotide Primers in Polymerase Chain Reactions and RT-PCR, Site- directed Mutagenesis and Cloning Vectors were explained, as well as the Application of Gene Expression Systems, Bioinformatics and Database Searching Microarray Technology.
The practical and skills training reinforced the lectures and provided essential skills in the current methodologies covering:
• Cleavage and Ligation of plasmid DNA,
• Transformation of bacterial cells,
• Preparation and Purification of Plasmid DNA
• Restriction Enzyme Mapping
• Agarose Gel Electrophoresis
• Recovery of DNA from Agarose Gels
• Preparation of Probes
• DIG-based Southern Blotting and Hybridisation
• DIG-based Colony Hybridisation
• Induction of Fusion Protein Production
• Polyacrylamide Gel Electrophoresis and Western Blotting
• Nucleic Acid Extraction
• Reverse Transcription and PCR
• DNA Sequencing and Computer-aided Analysis
Upon return to the research lab at the Heart Research Institute, I am now well equipped with new knowledge and expertise which will be invaluable in the process of designing new experiments and would like to thank ASI for their support.
