Molecular Biology Methods

Abstract

This handout is a complete course in Molecular Biology Methods for 3rd-year Biotechnology / Plant Genomics students. It covers the structure and properties of DNA and RNA, methods for DNA extraction and purification, and the main enzymes used in genetic engineering (restriction endonucleases, ligases, CRISPR-Cas systems) and their applications (cloning, mapping, fingerprinting, site-directed mutagenesis, recombinant DNA).

It then describes electrophoresis and hybridization techniques (Southern and Northern blot), PCR and its variants, mutagenesis approaches (including transposon-based methods), vector cloning, construction of genomic and cDNA libraries, and the basic principles of Sanger and next-generation sequencing. Finally, it introduces essential sequence bioinformatics tools and database searches (e.g. NCBI), providing both theoretical foundations and practical guidelines for applying molecular biology techniques in the lab.