HOME DETERGENTS AND DNA EXTRACTION FROM FRUIT Dana Hogan Problem • Testing which home detergent- shampoo, laundry detergent, or dishwashing detergent- most effectively extracts DNA from kiwi Research • DNA, or deoxyribonucleic acid, is the genetic material for all • • • • • living things that can be found in the cell nucleus. DNA extraction is the removal of deoxyribonucleic acid from the cells or viruses in which it can normally be found. In a basic DNA extraction, the steps included: lyses of tissue and/or cells, precipitating the DNA, the application of a detergent, and heat denaturation. The kiwifruit, or Actinidia deliciosa, is made-up mainly of sugar and ascorbic acid. A surfactant, a main component of cleaning detergents, is a substance that has the ability to remove dirt, grease, or oil from other surfaces when it is added to water. Previous student studies on the subject of the effectiveness of home detergents and DNA extraction from fruit found that laundry detergent proved the most effective. Hypothesis • If the effectiveness of shampoo, laundry detergent, and dishwashing detergent in DNA extraction from fruit were tested, then dishwashing detergent would prove the most effective. Materials • • • • • • • • • • • • • pH paper • 200mL cold ethanol • Volumetric flask • Freezer • Balance weighing to .01g • Balance weighing to • .001g • Small blender Fork and knife • 480g mashed kiwi • 20mL Suave® shampoo • 20mL all® laundry • detergent • 20mL Seventh • Generation™ dishwashing detergent • 360mL distilled water • 8g salt 500mL beaker 100mL graduated cylinder 6 200mL beakers Hot water bath Thermometer 2 large Tupperware containers Ice Cheesecloth Coffee filters 6 1000mL beakers Filter Strainer 10mL graduated cylinder 20 test tubes • Glass stirring rod • Small metal hook • Weighing paper Procedure • PH balance of detergents was tested. • A salt-water solution of 180mL of water and 4g of salt was prepared. • 60g mashed kiwi were poured into a 200mL beaker and researcher 10mL • • • • • • • • • • detergent and 30g salt-water solution were added. Mixture heated in hot water bath at 50ºC for 10 minutes, stirring occasionally. Mixture transferred to ice bath for 5 minutes. Cooled mixture was filtered into 1000ml beaker. (In first set of trials, cheesecloth was used for filtration, whereas in the second set of trials, coffee filters were used for filtration.) Once 15mL of filtrated were gathered, 3 test tubes with 5mL of filtrate each were filled. 10mL of cold ethanol were carefully poured into each test tube. Test tubes placed in ice bath for 5 minutes. Spooling of visible DNA was attempted. Spooled DNA was spread onto pre-weighed weighing paper and let dry for 2 days. Mass of DNA extracted was calculated. This procedure was repeated twice for shampoo and dishwashing detergent samples. In the first set of trials, 1 control sample was tested, but in the second set of trials, 3 control samples were tested. In first set of trials, 3 samples of laundry detergent were tested, but in the second set, 1 sample was tested. Variables • Independent variable- type of detergent added • Dependent variable- amount of DNA extracted • Control- samples without detergent • Constants- amount of salt-water, kiwi, and detergent added, temperature during incubation, length of time allotted for incubation and cooling, amount of filtrate used, amount of ethanol added to filtrate Data DNA Extracted from Each Sample (g) Control Shampoo Laundry Dish (1) Dish (2) Trail 1 0 0 0 0.01 0.124 Trial 2 0 0 0 0.02 0.099 Trial 3 0 0 0 0 0.095 Average 0 0 0 0.01 0.106 Data Average of DNA Extracted (g) Average DNA Collected (g) 0.14 0.12 0.1 0.08 0.06 0.04 0.02 0 Control Shampoo Laundry Detergent Dish (1) Dish (2) Photos Conclusion • The purpose of this experiment was to see which home detergent- shampoo, laundry detergent, or dishwashing detergent- would most effectively extract DNA from fruit. • The hypothesis that if the effectiveness of shampoo, laundry detergent, and dishwashing detergent in DNA extraction from fruit were tested, then dishwashing detergent would prove the most effective was supported. • Knowledge gained through this experiment can help find more cost effective methods of DNA extraction for various purposes, whether it is in schools or advanced laboratories. Discussion • Possible errors that occurred during experimentation include different filtration methods, varying methods for mashing the kiwi, and the temperature of the ethanol. • If repeated, more samples would be tested, coffee filters would consistently be used as the method of filtration, a balance measuring to .001 would be consistently used, and the ethanol would remain in the freezer up until it was needed. • Further experimentation could include testing different types of fruits and organic vs. nonorganic detergents. References • Bachor, K. (n.d.). The best detergent for plentiful DNA extraction. Retrieved from • • • • • • • • • • http://www.odec.ca/projects/2006/bach6k2/index.htm Berlow, L. H. (2011). Laundry detergent. Retrieved from eNotes.com, Inc. website: http://www.enotes.com/how-products-encyclopedia/laundry-detergent Morton, J. F. (1987). Fruits of warm climates (pp. 293-300). Retrieved from http://www.hort.purdue.edu/newcrop/morton/kiwifruit_ars.html Ombrello, T. (n.d.). The Pineapple. Retrieved from http://faculty.ucc.edu/biologyombrello/pow/pineapple.htm Rice, G. (2010, October 28). DNA extraction. Retrieved from http://serc.carleton.edu/microbelife/research_methods/genomics/dnaext.html Strawberry. (2011). In Britannica. Retrieved from http://www.britannica.com/EBchecked/topic/568585/strawberry Surfactants. (2005). Retrieved from Procter&Gamble website: http://www.scienceinthebox.com/en_UK/glossary/surfactants_en.html Understanding automatic dishwashing. (2010). Retrieved from American Cleaning Institute website: http://www.cleaninginstitute.org/clean_living/understanding_automatic_dishwashing.aspx#De tergent What is DNA? (2011, October 4). Retrieved from US National Library of Medicine website: http://ghr.nlm.nih.gov/handbook/basics/dna What is hair shampoo? (2011). Retrieved from Herbal Luxuries Natural Skin Care & Acne Products website: http://www.herballuxuries.com/what-is-hair-shampoo.html World of Forensic Science. (2005). DNA isolation methods. Retrieved from Encylcopedia.com website: http://www.encyclopedia.com/topic/DNA_Isolation_Methods.aspx