24 th oy CORE EXPERIMENT—8 pNolt | A1solation of DNA from Plant Materials % INTRODUCTION }, —— | Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) are the two types of nucleic \ acids found in living systems. DNA acts as the genetic material in most of the organisms. RNA, though, it also acts as a genetic material in some viruses, mostly functions as a messenger | adapter, structural and in some cases as a catalytic molecule. | ‘All human knowledge, especially of natural sciences is directed to develop technologies .\ for the comfort and well being of human beings Biotechnology has emerged as an off shoot of | modern biology in the twentieth century. The current break throughs in the field of biotechnol- | ogy are production of genetically modified organisms (plants, animals and micro organisms) | through recombinant DNA (r DNA) technology. Recombinant DNA technology (Genetic engi- | neering, has allowed breeders to introduce foreign DNA in other organisms, including bacteria, | yeasts, animals and plants). Such organisms are called Genetically Modified Organisms (GMOs). \ Thus, r DNA technology involves isolation of DNA from a variety of sources and formation of “ new combination of DNA. EXPERIMENT 8.1 throw bern Objective. To isolate DNA from available plant material such as spinaeheaves, REQUIREMENTS Plant material (such as spi a pestle, beakers, test {tubes eng engymes elu, protease, ribonuclease, lipases), ethanol, spool ete: Solt, gute) cle Aexqen} PROCEDURE Take a small amount of plant material (such-as-spinach_leaves/green-pea-seeds/green “ papaya) and grind it in a mortar. * Break the cell wall and other envelope of plant cell, by treating the material by enzyme cellulase. © ‘Treat the material with enzyme protease to remove histone proteins which are inter- twined with DNA. * Dissolve the associated RNA with enzyme ribonuclease. * Also dissolve lipids with enzyme lipase. © Precipitate out the DNA by adding chilled ethanol. 54— CORE EXPERIMENTS ss a _esnrioeesevartrmm Separate out the precipitated DNA as fine threads by spooling i.e., winding of the fine threads of DNA on real. Fig. 8.1, DNA that separates out can be removed by spooling (spool = reel for winding yarn). (OBSERVATION DNA appears as white precipitate of very fine threads on the spool. PRECAUTIONS 1. All the glasswares used must be thoroughly cleaned and dried. 2. The enzyme and other chemicals used for the experiment must be of standard quality. VIVA VOCE Q. 1» What is biotechnology ? Ans. Biotechnology deals with techniques of using live organisms or enzymes from organ- isms to produce products and processes useful to humans. Q. 2. What is recombinant DNA ? Ans. Recombinant DNA is the DNA formed by combining DNAs from two different sources! organisms. Q. 3. What is genetic engineering ? Ans. If refers to the techniques to alter the chemistry of genetic material (DNA or RNA) to introduce these into host organisms and thus change the phenotype of the host organisms. Q. 4. What are Genetically Modified Organisms (GMOs) ? Ans. These are the organisms whose genes have been altered by manipulation, Q. 5. Why is the enzyme cellulase used for isolating genetic material from plant cells ? Ans. The enzyme cellulase is used to digest the cellulosic cell wall present in plant cells.