‘om Isolation of P wal are the two types of Tug RNA) rrr in most of the organisms j° poe ; INTRODUCTION asonucleic ci , 1d ribonve’ ig materi ‘i i Deoxyribonucleic acid eed as the genetic M2. mpostly functions as a Messen 7 5 DNA acts a8 © ie acids found in living systems." vial in some ule. though, it also acts as a genetic materi? catalytic molect M girected 10 develop technol sages srr and in same cS ens HO emerged 88 an off “te Sci atu jotechndl : B sho, All human knowledge, espe ings Biotin a ughs in the field of biota for he eomfort and well ing ofA Trent break OO eal and micro organ modern biology in the twentie Mig modified or anisms (plant * DNA technology (Genetic mx oxy are production of genetiony AO nology. Recomm organisms, including bata r er other organisms, includ cor Seal besa to introduce foreign p 2 A anetically Modified Organisms (Gyo, ae aye plants): Such organisms arecall@d etary of sourees and formation, Thus, r DNA technology involves isolati new combination of DNA. ion of DNA fr Objective. To isolate DNA from available plant material such as spinach leaves, green pea seeds, papaya etc. REQUIREMENTS Plant material (such as spinach leaves, green pea seeds or green papaya), mortar and pestle, beakers, test tubes, liquid detergent, non-iodised sodium chloride, distilled water, meat tenderizer or papain solution/juice of papaya/pine apple juice, 95% ethanol, spool etc. PREPARATION OF SOLUTIONS + Detergent salt solution is prepared by adding 10 mL liquid detergent and 10 g of non- iodised sodium chloride to 90 mL. of distilled water. Meat tenderizer solution is prepared by addi q died water ie papel spl iad hrngh meal cates bese 8% NaCl solotion is prepared by dissolving 6 g of no Chilling of ethanol must be done by keeping 95% ethanol in plastic bottle in the freeze" over night. n-iodised sodium chloride in 100 54= URE FqoceDuRe . aya) and grind it in Peake 5K of the plant tissue (spinach leaf/ : green pea seed/green pap: the mortar by adding 10 mL detergent, salt solution and filter it through muslin cloth. ‘take 10 mL of the filtrate, add 3-4 mL tenderizer/ ice and swirl th . 2 papaya juice and swin! by holding the tube between the two hands to mix the contents. |, Pour 10 mL chilled 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. |, Using the glass rod stir gently through interface of the two layers to collect the precipi- tate of DNA and place it in a test tube with 5% NaCl or distilled water. | The quantity of DNA present in the given plant material can be estimated through spectrophotometer. e test tube Fig. 8.1. 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. PRECAUTIONS 1. 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 be manufactured by standard pharmaceuticals. ee