| Isolation of DNA from Plant Materials INTRODUCTION Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) are the two types of nucleic acids found in living eee DNA acts as the genetic material in most of the organisms. RNA, though, it also acts asa 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 biotechnology are production of genetically modified organisms (plants, animals and micro organisms) through recombinant DNA (r DNA) technology. Recombinant DNA technology (Genetic engineering, has allowed breeders to introduce foreign DNA in other organisms, including bacteria, yeasts, animals and plants). Such orgenisms ae called Genetically Modified Organisms (6MOs). Thus, r DNA technology involves isclation of DNA from a variety of sources and formation of new combination of DNA, (EXPERIMENT 8.1 ie isolate DNA from available plant material such as spinach leaves, green pea seeds, papaya etc, «~-REQUIREMENTS vo beakers, test tubes, liquid detergent, non-iodised sodium chloride, Papain solution juice of papaya/pine apple juice, 95% ethanol, spool etc, PREPARATION OF SOLUTIONS + Detergent salt solution is prepared by addin, sodium chloride to 90 mL of distilled water, * Meat tenderizer solution is prepared i i is Preps ved by adding 5 g. of tenderizer (enzyme) to 95 mL. of distilled water (Juice of papaya/pine apple, filt : pear yanl . paya/Pine apple, filtered through muslin, cloth can be used as substitute for 1 10 mL liquid detergent and 10 g of non-iodised distilled water. ng 95% ethanol in plastic bottle in the freezer over * Chilling of ethanol must be done by keep night.aye Experiments é | procEDURE + Take LOmL of the filtrate, add 3-4 mb te Take 5 g of the plant tissue (spinach leaf/green pea seed/green papaya) and grind it in the mortar by adling 10 mb, detergent, salt solution and filter through muslin cloth nderizer/papaya juice and swirl the test tube by holding the tube between the two hands to mix the content Pour 10 mL chilled ethanol care! at hilled ethanol carefully down the side of test tube to form a layer on the top of the content; let it stand undisturbed for about 3 minutes, J: ‘od stir ger i i Us rod sti gently through interface of the two layers to collect the precipitate of DNA and place it ina test tube with 5% NaCl or ng the gl illed water. The quantity of DNA present in the given plant material can be estimated through spectrophotometer DNA that separates out can be removed by spooling (spool = reel for winding yarn). OBSERVATION, ~ ‘The addition of ethanol to the solution causes DNA to precipitation. The DNA fibres appears as white precipitate of very fine threads on the glass spool. PRECAUTIONSL~ Z ‘The plant material should be washed throughly with distilled water to remove any dust and dried by blotting before weighing. All the glasswares used must be thoroughly cleaned and dried. ‘The chemicals and enzymes used for the experiment must be of standard quality which should mii be manufactured by standard pharmaceuticals.