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    BIOTECH Project, University of Arizona DNA Extraction From Kiwifruit

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    Yuliana Purnamasari
    1) DNA was extracted from kiwifruit to allow students to purify and observe DNA. 2) A solution containing soap and salt was used to break open the kiwifruit cells and dissolve membranes while precipitating proteins and carbohydrates. 3) The mixture was cooled and mushed repeatedly to protect the DNA from enzymes while allowing cell contents to be released. Most cell parts were filtered out while DNA remained in solution.

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    BIOTECH Project, University of Arizona DNA Extraction From Kiwifruit

    Uploaded by

    Yuliana Purnamasari
    39 views2 pages
    1) DNA was extracted from kiwifruit to allow students to purify and observe DNA. 2) A solution containing soap and salt was used to break open the kiwifruit cells and dissolve membranes while precipitating proteins and carbohydrates. 3) The mixture was cooled and mushed repeatedly to protect the DNA from enzymes while allowing cell contents to be released. Most cell parts were filtered out while DNA remained in solution.

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    1) DNA was extracted from kiwifruit to allow students to purify and observe DNA. 2) A solution containing soap and salt was used to break open the kiwifruit cells and dissolve membranes while precipitating proteins and carbohydrates. 3) The mixture was cooled and mushed repeatedly to protect the DNA from enzymes while allowing cell contents to be released. Most cell parts were filtered out while DNA remained in solution.

    Copyright:

    © All Rights Reserved
    Download as DOC, PDF, TXT or read online from Scribd
    Download as doc, pdf, or txt
    39 views2 pages

    BIOTECH Project, University of Arizona DNA Extraction From Kiwifruit

    Uploaded by

    Yuliana Purnamasari
    1) DNA was extracted from kiwifruit to allow students to purify and observe DNA. 2) A solution containing soap and salt was used to break open the kiwifruit cells and dissolve membranes while precipitating proteins and carbohydrates. 3) The mixture was cooled and mushed repeatedly to protect the DNA from enzymes while allowing cell contents to be released. Most cell parts were filtered out while DNA remained in solution.

    Copyright:

    © All Rights Reserved
    Download as DOC, PDF, TXT or read online from Scribd
    Download as doc, pdf, or txt
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    BIOTECHProject,UniversityofArizona

    DNAExtractionfromKiwifruit

    Teacher Guide: What is DNA? DNA Extraction from Kiwifruit


    Adapted from the Office of Biotechnology, Iowa State University
    Introduction
    DNA is present in the cells of all living organisms. This procedure is designed to extract DNA from kiwi in
    sufficient quantity to be seen and spooled. This activity is ideal for students to work in pairs, but each student
    will have a tube of DNA at the end.
    Some questions to get you thinking about todays lab:
    One way to purify a molecule is to get rid of everything but that molecule. If we want to isolate DNA from
    kiwifruit, what do we have to get rid of?
    All parts of the cell besides the DNA, i.e. cell wall (kiwi is a plant, after all), cell membrane, mitochondria,
    Golgi apparatus, endoplasmic reticulum, vacuoles, lysosomes, nuclear membrane, etc.
    What materials would you use to do that?
    Something to mush the cells (blender or your hands), something to destroy membranes (soap dissolves
    them), something to get rid of proteins and carbohydrates (salt causes them to precipitate), something to
    separate insoluble cell stuff from soluble DNA, and something to help get the DNA (alcohol precipitates
    it).
    What can we do with the DNA once weve purified it?
    Use it in DNA fingerprinting (solve a crime, see a genetic defect), put it into another organism to give it
    specific traits (this is called transformation or genetic engineering), other?
    Materials

    ziplock bags (1 per student pair)


    jar or beaker that fits strainer or funnel (1 per 8 students)
    strainer or funnel (1 per 8 students)
    cheese cloth (cut to cover the funnel)
    ice water bath (a large mixing bowl works well) (1 per 4-8 students)
    extraction solution (1 tube of 20 mls per student pair)
    kiwifruit (cut into 12-16 pieces, each student pair needs 4 pieces)
    cold 95% ethanol or isopropanol (2-3 mls per student)
    small test tubes (1 per student)

    Extraction solution recipe:


    For one liter of the extraction solution, mix 100 ml of shampoo (eg Suave Daily Clarifying Shampoo, many
    shampoos will work, but do not use shampoos with conditioner or baby shampoo) and 15 g of table salt (iodized or
    non-iodized both will work). Add water to make a final volume of 1 liter. Dissolve the salt by stirring slowly to
    avoid foaming. Measure 20 ml of solution for each pair of students.
    Protocol
    1. Get 4 pieces of kiwi and put them in a ziplock bag.
    2. Add 20 ml of extraction solution to the ziplock bag. Make sure the bag is closed without much extra air.
    Mush the kiwi thoroughly but carefully so the bag doesnt break, for about 5 minutes. What does mushing the
    kiwi do?
    Breaks the cell wall
    3. What do you think the extraction solution is? What does it do to the kiwi?

    BIOTECHProject,UniversityofArizona
    DNAExtractionfromKiwifruit

    Soap will cause the solution to bubble so students should be able to guess what's in here. The soap
    destroys the cell and nuclear membranes, allowing the DNA to get out. There is also salt in the extraction
    solution, which causes the proteins and carbohydrates to precipitate, while the DNA remains in solution.
    4. Cool the kiwi mixture in the ice bath for a minute. Then mush the kiwi more. Cool, then mush. Repeat this
    several times. Why do we cool the mixture?
    Cooling protects the DNA. There are DNases (enzymes that destroy DNA) in the cell's cytoplasm. The
    DNA is usually protected from DNases by the nuclear membrane, but that is destroyed by the soap.
    Cooling slows down the DNases, just like it would any enzymatic reaction. DNases are in our cells to
    protect us from foreign DNA (like viruses).
    5. Filter the mixture through the cheesecloth. All the groups can combine their mixtures at this point, to filter
    together. What is being filtered out? What is going through the filter?
    Students can usually see the seeds being filtered out. Most of the cell parts and the precipitated protein
    and carbohydrate are also being filtered out at this point.
    6. Dispense approximately 2 ml of kiwi solution into each test tube, one for each pair.
    7. Being careful not to shake the tubes, add approximately 2 ml of cold 95% ethanol to each tube. What do
    you think the ethanol does? Why do we want it cold?
    We don't have to worry about the DNases at this point, because hopefully they've mostly been filtered
    out. What we are most concerned about is precipitating (or solidifying) the DNA. The colder something
    is, the more likely it will precipitate or solidify. Cooling the alcohol just increases the amount of DNA
    that precipitates.
    8. Take a look at your tube. What do you see in the top portion of the liquid?
    You can actually pick up the DNA at this point, using a toothpick, wood pencil, or glass stirring rod.

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