The field of molecular biology has a profound impact in life science investigation. The development of more and more sophisticated experimental techniques in molecular biology with a broad, interdisciplinary applicability have stimulated research and progress in almost all the disciplines of life science. The rationality and approach of scientific experimentations have changed and allowing revolutionizing discoveries not only in molecular biology but also in biochemistry, biophysics, biotechnology, cell biology, and genetics. Reserachers can choose suitable techniques based on their usefulness, objectives and the financial and temporal resources available to the them. Many molecular biology techniques have extensively been used in the study of gene expression.
Gene expression is the process by which information from a gene is used in the synthesis of a functional gene product (mRNA) that enables it to produce end products (protein), and ultimately affect a phenotype, as the final effect.
The techniques for mRNA expression analysis are very sensitive, speedy and provide complete picture. The study of gene expression involves the comparison of mRNA populations between two samples, treated versus untreated, diseased versus healthy, stage A of development versus stage B of development. The techniques used to study mRNA expression are generally divied into hybridization-based techniques, PCR-based techniques and sequencing-based techniques.
Hybridization-based techniques employ hybridization process to detect particular sequences within a complex mixture of DNA or RNA molecules. The three most popular hybridization based techniques i.e., Northern blot, RNA-Fluorescence in situ hybridization (RNA-FISH) and DNA microarray are discussed in this course in detail.
PCR-based techniques exploit the ability of Taq polymerase from Thermus Aquaticus to amplify DNA by using oligonucleotides (primers) that are a complementary sequence to the target DNA region. Reverse transcriptase polymerase chain reaction (RT-PCR) and real time qRT-PCR fell under this category and elaborated in this course.
Sequencing-based techniques determine the exact nucleotide (A, C, G, and T) order of a given DNA fragment by using sanger method or high throughput sequencing/next generation sequencing technology. In this course, serial analysis of gene expression (SAGE) and RNA sequencing (RNA seq) are explained.
Cancer can be described as a disease of altered gene expression. It is caused by the mutations in the genes that control the way our cells function, especially how they grow and divide. Scientists are working to understand the common changes that give rise to certain types of cancer or how a modification might be exploited to destroy a tumor cell.
Oncogenes and tumor suppressor genes control the normal cell growth and any mutation in these genes lead to the development of cancer. The difference between normal cell and cancerous cell pathways can be studied by analyzing gene expression levels and the biological pathways associated with the genes involved in a cancer. Lab techniques and web servers are developed by scientists to study and compare results with the already available data on different cancers.
Targeted therapy for cancer have exploited the overexpression of a specific protein or the mutation of a gene to develop a new medication to treat disease and has given rise to the field of personalized medicine.
This course is a valuable resource for students and researchers related to molecular biology, forensic science, medical laboratory technology, biotechnology, and genetics.
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