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    Lec-Bio103L, Expt 7 SUMMER 2024

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    Lec-Bio103L, Expt 7 SUMMER 2024

    Uploaded by

    mustakimislamt
    1 views11 pages

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    Copyright:

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    1 views11 pages

    Lec-Bio103L, Expt 7 SUMMER 2024

    Uploaded by

    mustakimislamt

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 11

    BIO103 Lab

    (1 credit hour)
    SUMMER 2024

    Dept. of Biochemistry & Microbiology


    Experiment 7
    Extraction of plant genomic DNA.
    Isolation of plant genomic DNA from spinach

    • Deoxyribonucleic acid (DNA) is a nucleic acid that contains the genetic instructions for
    the development and function of living things.

    • The structure of DNA is called a double helix, which looks like a twisted staircase.

    • The main role of DNA in the cell is the long term storage of information's.

    • There are four different bases:

    □adenine (A), cytosine (C), guanine (G), or thymine (T).

    • The two strands are held together by hydrogen bonds between the bases, with adenine
    forming a base pair with thymine, and cytosine forming a base pair with guanine.
    Structure of DNA
    Structure of DNA

    5
    Functions of DNA

    DNA has many important functions in living systems, such as-


    • DNA contains the instructions needed for an organism to develop, survive and
    reproduce.
    • It stores instructions for making other large molecules, called proteins.
    • It is the genetic material passed from parent to offspring.
    • Diagnosis of Genetic disorder.
    • Forensic Test for criminal investigation, paternity testing.
    • Gene therapy works by replacing or inactivating disease-causing genes

    7
    DNA Extraction
    • DNA can be isolated from living tissues e.g., spinach, banana, onion.

    • DNA isolated from the nucleus, for which cell wall and plasma membrane must be broken
    down by mechanical forces, in the presence of salt and detergent.

    • Isolation of DNA from plant material poses special challenges, i.e., several plant
    metabolites have chemical properties similar to DNA.

    • Young and healthy tissues are recommended to use for DNA extraction

    □ Young tissues contains more cells than older tissue.

    □ Young tissue of the same weight contains fewer metabolites.


    Principle

    • The process of isolating nucleic acid is a crude extraction method.

    • By using salt and detergent solution causes the cell membrane to break down or
    lyse and emulsifies the lipids and proteins of the cell.

    • After incubation period, the crude mixture is filtered to get homogenized solution.

    • Pouring of this solution into petri dish and glass tube containing 95% ethyl alcohol
    or ethanol, causes the precipitation of DNA in time.

    • After some moment, it appears as thick, stringy, white or slight greening mass.

    • As DNA is not soluble in alcohol, can be isolated as they clamp together and
    become visible to the naked eye.
    EXPERIMENT
    • Materials • Procedure

    • Measuring cylinder,
    beakers Chop and grind sample

    • Mortal and pestle

    • Light colored dish Add liquid with detergent and salt


    solution
    washing liquid

    • Ethanol 95%

    • Large spoon for mixing Incubate at 56°C for 10 minutes

    • Petri dishes

    • Strainer/ funnel Filtration

    • Salt

    • Filter cloth or paper Add 95% ethanol and visualize DNA


    Thank you!

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