Introduction to Molecular Biology Techniques

This is a three-day course for those interested in learning both fundamental and advance level molecular biology techniques. Molecular biology techniques are being used extensively in modern day drug discovery, research and development, and diagnostics. It is intended for personnel with some scientific background seeking basic and advance level molecular biology training. The course should be informative for personnel involved in, or participating in, molecular biology related and biotechnological research, supervision, scale-up or manufacturing and other technical operations.

Learning Objectives

Upon completion of this course, you will have gained an understanding of techniques that are currently being utilized in the biotechnology and pharmaceutical industries. You will be familiar with concepts pertaining to basic molecular biology principles and techniques for understanding various contemporary areas of research and their applications. You will have acquired the facility for communication with associates in various areas of molecular biology.

Course Description

Molecular biology pertains to the study of living systems at the molecular level and provides a background appropriate for further work in the growing technologies of genomics, cell biology, biotechnology, microbiology, diagnostics and therapeutics. This course will focus on selected aspects of molecular biology that provide the non-specialist with the principles for understanding the structure and function relationships of molecular biology techniques including DNA manipulation, sequencing, cloning, subcloning, library construction, screening, RNA isolation and characterization, analysis of expression, microarrays, gene chips and RT-PCR. Multiple modern day molecular biology techniques in biotechnology and pharmaceutical industry will be highlighted. Several examples of molecular techniques will be demonstrated in case studies and examples.

________________________________________

COURSE AGENDA

DAY ONE

Introduction - Principles of Molecular Biology Techniques

Introduction to asics and advance information on DNA, RNA and roteins. Potential applications of molecular biology techniques in drug discovery and development will also be reviewed

DNA Molecular Techniques

Isolation and purification of DNA samples from different cell types and tissues, DNA concentration techniques, estriction digestion and analysis (Southern blot analysis), el electrophoresis, labeling of DNA, andom subclone generation, andom fragment end-repair, size selection, phosphorylation, DNA ligation, ompetent cell preparation and acterial cell transformation.

DNA Library construction

Fundamentals of DNA library construction, detail analysis of the protocols, enzymes and DNA utilized, steps involved, titering the phage library, screening the library, identification and characterization of clones.

DNA sequencing

Basics and applied methods of DNA sequencing, modern day tools and instruments for sequencing, Applications of sequencing in drug discovery and development, SNPs identification and characterization techniques, SNPs applications in modern day drug application.

RNA Technologies

RNA Isolation and Characterization

Isolation, purification and characterization of total and PolyA+ RNA from cells and tissues using molecular techniques, reverse transcription and cloning.

DAY TWO

PCR (Polymerase Chain Reaction)

Fundamentals of PCR, PCR amplification tools and technologies, PCR cloning and applications of PCR technique.

RT-PCR

1st strand cDNA synthesis and PCR. Quantitative RT-PCR of cDNA samples with TaqMan, cloning and characterization.

Differential Display PCR

Identified and characterized differentially expressed novel genes (from tumor and other tissues) and under different biological conditions using DD-PCR technique, case studies and applications.

Northern Blotting

Blotting of isolated and purified total and/or polyA+mRNA from several cell types and tissues. Detection using radioactive and non-radioactive DNA, RNA and oligonucleotide probes.

Reverse Northern

Experience in performing reverse Northern using Dot and Slot blot apparatuses.

In Situ Hybridization

Fundamentals of in situ hybridization technique. Protocol overview of gene expression analysis in tissues.
Phage library construction and screening

Tools and technologies in constructing a phage library. Subtractive library construction and screening: Detail analysis of the protocols, enzymes and RNA utilized, steps involved, titering the phage library, screening the library, identification and characterization of clones and their sub-cloning. Differential library hybridization and differential library screening of libraries for expression analysis.

DAY THREE

RNase Protection Assay (RPA)

RPA fundamentals and experimentation details, Applications (quantitative differential expression of genes in different tissues, cells or under different growth conditions).

RNAi

Fundamentals of RNAi and siRNA techniques and applications in modern day drug discovery

Differential Display PCR

Basics and advance information on the technique, protocols, applications and case studies in isolating differentially expressed genes.

SAGE (Sequential Analysis of Gene Expression)

Fundamentals of SAGE technology, SAGE library construction, SAGE tags, SAGE library sequencing, application in drug discovery and development.

Macroarrays

Basics of macroarray technology. Surfaces and protocols for spotting nucleic acids, formats and designs. Identification of differentially expressed genes by differential hybridization of macroarrays.

Microarrays

Fundamentals of microarray technologies. Protocols to design and develop microarrays.
Identification of differentially expressed genes by differential hybridization of arrays of low and high density from commercial sources.

Gene Chips

Basic information on the synthesis, design and applications of gene chip technology. Applications in drug discovery and development.

Protein Technologies

Isolation and characterization of proteins. Introduction to proteomics techniques.

Applications of DNA and RNA technologies in Drug Discovery and Development