Chemistry of DNA extraction核酸提取的化学.ppt

Detergent/Enzyme/Heat Break down the cell wall 60o C breaks down nuclear membrane, but not DNA; 80o C will break down DNA as well DNA Extraction Procedure - 1 Blend for 15 seconds: DNA source – 100 ml green split peas Cold water – 200 ml Large pinch of table salt Pour mixture through a strainer Add detergent – 2 tablespoons Let sit for 5 – 10 minutes DNA Extraction Procedure - 2 Pour mixture into test tubes – 1/3 full Add meat tenderizer – dip of wet stick Tilt test tube Slowly pour rubbing alcohol into the tube Equal amounts of alcohol DNA mixture Watch DNA appear in the alcohol Salt (Na +) is attracted to the phosphate group Denatures the protein that lyses (breaks down) DNA R-OH Water is HOH, like dissolves like Alcohol dissolves the water and pulls it away from the DNA R is shorthand for any organic group or hydrocarbon * Chemistry of DNA extraction - some bio too! DNA Extraction from Green Split Peas or raw wheat germ! DNA appears at alcohol/ mixture interface and can be spooled onto a stirring rod Alcohol Cold for two reasons: Slows down enzymes that break up DNA DNA is less soluble in cold temperatures Where can you find DNA? In the cells of any living thing. The only cells that do not have DNA are red blood cells. In the extraction procedure we use a blender to separate the cells from one another. The cell is surrounded by a sack (cell membrane). Where is DNA located in the cell? In the nucleus. (There is also some DNA in the mitochondria.) Notice in the diagram that the nucleus of the cell is also surrounded by a membrane sack. To see the DNA we have to break open these sacks. We can do this with detergent. How can we use detergent to break open the membrane? Think about why you use soap to wash dishes or your hands. To remove grease and dirt, right? Soap and grease molecules are made of two parts: Heads, which like water And tails which HATE water Both soap and grease molecules organize themselves into bubbles when they are in water. This is