De extracted from any living thing. DNA in the nucleus which is responsible for maintenance and inheritance of characters. Object: To isolate DNA from plant materials like spinach, leaves, green pea seeds, papaya. Requirements: Spinach leaves, green pea seeds, papaya (5-10 g), grinder, crushed dry ice, CTAB - buffer solution (25%), chloroform: isoamyl alcohol 24 : 1), ice-cold isopropanol, 5 M NaCl, oven, water bath and centrifuge etc. Procedure 1, Weigh 5 g of plant tissue (spinach/green pea seeds etc). 2. Quickly freeze the plant tissue in liquid nitrogen. 3, Grind the frozen material with dry ice in a grinder. 44 Transfer the material into centrifuge tube and add 3 ml of extraction buffer (CTAB buffer solution). Mix it by inversion to form a slurry. 5, Water bath the material at 60°C for 30-60 minutes. 6 Add 3 ml of chloroform: isoamyl alcohol (24: 1) and mix the contents by inversion for about 10-15 > minutes. 7, Centrifuge the contents at 8000 rpm for about 10 min. §, Transfer the supernatant liquid (upper aqueous layer which has DNA) to another 10 ml centrifuge tube. 9. Add 1.5ml of 5 M of NaCl and mix properly the contents. 10. Add ice-cold 0.6 volume of isopropanol. |. Careful mixing the mixture (after 60 minutes) will produce fibrous DNA. It can be removed by spooling. amount of DNA is measured and estimate the DNA content with the help of spectrophotometer. with animal or human DNA. i like grapes which are watery because enough DNA will not be th conditions of the mixture. taken when liquid nitrogen is used. should be taken. ere to see.- Requirments: Plant material like banana, mortar and pestle, salt, liquid detergent (dish soap) p "water, measuring cylinder, contact lens cleaner or Pineapple juice, glass rod. beaker, pipett, test tube ete Take 100 g of banana & smash it by using mortar & pestle. * Take this paste of fruit in a beaker . * Add 2g salt and 5 mi of liquid detergent (dish soap) in 100 ml of water. * Gentle pour this solution to the beaker having squished fruit. Gently mix the solution. * Let this mixture settle down for 10-20 minutes. * Filter this mixture by using coffer filter. * Take 5 mloffiltered solution in a test tube and add Iml of pineapple juice or contact lens cleaner. | © Wait for 2-3 minutes * Now pour 70% ice cold ethanol down the side of the test tube. ‘* DNA appear in the test tube in the form of white strings * Collect the DNA with the help of glass rod. ‘Alcohol — Alcohol DNA Test tube Solution of DNA in test tube ipitate at the junction of solution & ethanol d be used.