Conventional and Basic Genomic Techniques

Abstract

AbstractIn this chapter, certain very basic and primary technologies pertaining to genomics have been recapitulated. The topics covered include basic protocols for genomic DNA isolation through various techniques, bacterial DNA isolation, Plasmid DNA isolation, and other electrophoretic techniques for its checking and visualization as agarose gel electrophoresis and Polyacrylamide gel electrophoresis. This chapter also includes certain basic polymorphism techniques suitable for detection of polymorphism, such as Restriction fragment length polymorphism (RFLP), Single stranded conformation polymorphism (SSCP). High-quality DNA is extremely essential for next generation sequencing studies, will be discussed in later chapters. The basic principle involved in genomic DNA isolation is the destruction of lipid and protein respectively through SDS and proteinase. DNA is precipitated through isopropanol. Plasmid DNA isolation involves alkali lysis which destroys bacterial DNA. In the electrophoretic techniques, DNA fragments are separated based on their mobility which depends on their fragment size. RFLP involves cutting of DNA fragments based on restriction enzymes, whereas SSCP involves differences in conformation of single stranded DNA due to their nucleotide variation and their mobility through PAGE.KeywordsDNA isolationElectrophoresisRFLPSSCPPAGE