Dna Extraction And Amplification

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Experiment No. 8 Extraction and Amplification of Rice Genomic DNA Introduction The intraspecific variation in Oryza sativa is remarkably extensive and subspecific classification has always been a matter of importance for rice breeders and geneticists. For a long time, rice has become divided into many varieties. Identification of these varieties can be acquired through molecular processes, with the help of their genetic markers. DNA Extraction is a process that is widely used in Cell molecular biology in isolating DNA from cells chemical, mechanical (grinding), or sonicating the sample breaking cells open and leaving the cell vulnerable for DNA extraction process. DNA extraction is also a routine procedure to collect DNA for subsequent molecular or forensic analysis; DNA sequencing and DNA fingerprinting. Refinements of the technique include adding a chelating agent to sequester divalent cations such as Mg2+ and Ca2+, which prevents enzymes like DNase from degrading the DNA. Cellular and histone proteins bound to the DNA can be removed either by adding a protease or by having precipitated the proteins with sodium or ammonium acetate, or extracted them with a phenol-chloroform mixture prior to the DNA-precipitation. DNA amplification is the cornerstone of modern biotechnology and it is also a key procedure in numerous basic studies involving DNA molecules. All methods for DNA amplification have rested on the concept of DNA strand complementarity discovered by James Watson and Francis Crick fifty years ago. To an equal extent, these methods became possible with the discovery of DNA polymerases first identified by Arthur Kornberg soon after the Watson-Crick discovery and DNA ligases discovered in 1967 by Martin Gellert, Charles Richardson, Jerard Hurwitz, Robert Lehman and others. Using these enzymes (and later their thermostable variants), a variety of

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