detergent/enzyme/heat break down the cell wall 60 o c breaks down nuclear membrane, but not dna; 80...
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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
Chemistry of DNA extraction - some bio too!
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DNA Extraction from DNA Extraction from Green Split Peas or raw Green Split Peas or raw wheat germwheat germ! !
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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
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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
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SoapGrease Grease and Soap
Heads
Tails
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DNA appears at alcohol/ mixture interface and can be spooled onto a stirring rod
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Salt• (Na +) is attracted to the phosphate
group• Denatures the protein that lyses
(breaks down) DNA
Alcohol
Cold for two reasons:
•Slows down enzymes that break up DNA
•DNA is less soluble in cold temperatures
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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