Molecular Clones V.II

Description

Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. The use of the word cloning refers to the fact that the method involves the replication of one molecule to produce a population of cells with identical DNA molecules. Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA. Molecular cloning methods are central to many contemporary areas of modern biology and medicine. Molecular cloning refers to the isolation of a DNA sequence from any species (often a gene), and its insertion into a vector for propagation, without alteration of the original DNA sequence. Once isolated, molecular clones can be used to generate many copies of the DNA for analysis of the gene sequence, and/or to express the resulting protein for the study or utilization of the protein's function. The clones can also be manipulated and mutated in vitro to alter the expression and function of the protein. Molecular cloning is a set of methods, which are used to insert recombinant DNA into a vector- a carrier of DNA molecules that will replicate recombinant DNA fragments in host organisms. The DNA fragment, which may be a gene, can be isolated from a prokaryotic or eukaryotic specimen. DNA cloning is the process of making multiple, identical copies of a particular piece of DNA. In a typical DNA cloning procedure, the gene or other DNA fragment of interest (perhaps a gene for a medically important human protein) is first inserted into a circular piece of DNA called a plasmid. The insertion is done using enzymes that "cut and paste" DNA, and it produces a molecule of recombinant DNA, or DNA assembled out of fragments from multiple sources. This book presents the principles of gene manipulation and its associated techniques in sufficient details and also covers biological replication of DNA, restriction and endonuclease and other related enzymes. Contents: 1. Molecular and Cellular Cloning, 2. Extraction, Purification and Analysis of DNA and RNA, 3. Bioinformatics, 4. Analysis and Manipulation of DNA and RNA, 5. Methods for in Vitro Mutagenesis, 6. Genes into Cultured Mammalian Cells, 7. Cloning, DNA and Genetic Engineering, 8. Quantification of DNA and RNA by Real-Time Polymerase Chain Reaction.