Author Description

This book has a distinguishing feature of having condensed material with adequate information on genetic engineering especially of the microbes. The book covers almost all the topics of genetic engineering for the graduate, postgraduate students and young research scholars of biological sciences. The book is written as per syllabus of genetic engineering paper for Masters course in biotechnology, biochemistry, life sciences of most of the universities. The book is much useful for the students of Masters degree. Emphasis is given on the basic fundamentals. The book contains twelve chapters starting from ' Isolation, purification and estimation of nucleic acids' as chapter 1. The chapter describes general techniques for the isolation and purification of DNA as well as RNA. It also describes methods for quantitative estimation of the nucleic acids. The second chapter describes general characteristics of the vectors used in genetic engineering and also the general account of commonly used individual vectors. The chapter also describes expression vectors. The third chapter describes various commonly used restriction endonucleases. The fourth chapter describes commonly used enzymes in genetic engineering viz. Reverse transcriptase, DNA polymerase I, polynucleotide kinase, teminal dcoxynucleotidyl transferase, alkaline phosphatase, SI nuclease, DNA ligase etc. The fifth chapter describes electrophoresis for the separation of nucleic acids fragments. The sixth chapter is of cloning strategies. It describes construction of genomic DNA library, chromosomal walking, cDNA library, cDNA cloning. The seventh chapter describes DNA sequencing techniques and includes chemical modification method of Maxam and Gilbert, dideoxy sequencing method of Sanger, modifications of chain terminator sequencing, analysis of the sequencing data. The eighth chapter includes various methods of site directed mutagenesis. The ninth chapter describes polymerase chain reaction (PCR). It also includes p