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    Lab3 Polytene Chromosome

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    Sohaib Omer Salih
    This document describes polytene chromosomes and a procedure for observing them under a microscope using Drosophila melanogaster larvae. Polytene chromosomes are giant chromosomes formed through repeated DNA replication without cell division in dipteran flies. This results in large banded chromosomes where the centromeres do not replicate well and bundle together. The document outlines how to dissect Drosophila larvae to remove and prepare the polytene-containing salivary glands for staining and microscopic observation of the large multistranded chromosomes.

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    Lab3 Polytene Chromosome

    Uploaded by

    Sohaib Omer Salih
    729 views17 pages
    This document describes polytene chromosomes and a procedure for observing them under a microscope using Drosophila melanogaster larvae. Polytene chromosomes are giant chromosomes formed through repeated DNA replication without cell division in dipteran flies. This results in large banded chromosomes where the centromeres do not replicate well and bundle together. The document outlines how to dissect Drosophila larvae to remove and prepare the polytene-containing salivary glands for staining and microscopic observation of the large multistranded chromosomes.

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    This document describes polytene chromosomes and a procedure for observing them under a microscope using Drosophila melanogaster larvae. Polytene chromosomes are giant chromosomes formed through repeated DNA replication without cell division in dipteran flies. This results in large banded chromosomes where the centromeres do not replicate well and bundle together. The document outlines how to dissect Drosophila larvae to remove and prepare the polytene-containing salivary glands for staining and microscopic observation of the large multistranded chromosomes.

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    729 views17 pages

    Lab3 Polytene Chromosome

    Uploaded by

    Sohaib Omer Salih
    This document describes polytene chromosomes and a procedure for observing them under a microscope using Drosophila melanogaster larvae. Polytene chromosomes are giant chromosomes formed through repeated DNA replication without cell division in dipteran flies. This results in large banded chromosomes where the centromeres do not replicate well and bundle together. The document outlines how to dissect Drosophila larvae to remove and prepare the polytene-containing salivary glands for staining and microscopic observation of the large multistranded chromosomes.

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    of 17

    Lab No ( 3 )

    Polytene Chromosomes

    Polytene chromosomes are giant chromosomes
    common to many dipteran (two-winged) flies.

    They begin as normal chromosomes, but through
    repeated rounds of DNA replication without any cell
    division (called endoreplication), they become large,
    banded chromosomes.

    For unknown reasons, the centromeric regions of the
    chromosomes do not endoreplicate very well. As a
    result, the centromeres of all the chromosomes bundle
    together in a mass called the chromocenter.






    The salivary gland cells in the larval stages of
    Drosophila contain large, multistranded polytene
    chromosomes.

    This laboratory procedure outlines the
    mechanism by which salivary glands are
    removed and prepared so that the polytene
    chromosomes may be observed.

    Drosophila melanogaster is easier to dissect and
    remove the salivary glands from the larvae of
    this species.
    Polytene
    Chromosomes
    from Salivary
    Glands
    Student Handout
    Materials
    For each student group:
    compound microscope
    2 teasing needles
    insect pins
    microscope slides
    cover glasses
    stock supply of Drosophila melanogaster larvae
    dropping bottle of aceto-orcein stain
    dropping bottle of 45% acetic acid solution
    Procedures

    Removing the salivary glands:
    1- Remove a large larva from the stock of D.
    melanogaster. Larger larvae are easier to
    dissect. However, select an active larva and one
    that has not started to pupate.
    2- Using the compound microscope, dissect the
    larva by placing one teasing needle on the
    posterior aspect of the larva and the other
    needle at the anterior end, near the black mouth
    parts.
    3- Carefully pull outward with the anterior needle.

    Procedure for removing Drosophila salivary glands
    ( cont )
    4- There are two transparent salivary glands
    located anteriorly in the larva.

    5- Discard all of the larva except for the
    salivary glands.
    Staining and Observing

    1- Place 2 drops of aceto-orcein stain on the
    salivary glands, and let it stand for 10 minutes.
    2- Place a cover slip over the glands, and using
    your thumb and a paper towel, push down on
    the slide. The pressure applied will squash the
    glands, rupture the nuclear membrane, and free
    the chromosomes.
    3- Using a compound microscope, observe the
    slide under low and high magnification.
    4- Make the slide permanent by brushing along the
    edges of the cover slip with clear nail polish

    The Puffing
    Puffing is the term that describes structural
    changes in polytene chromosomes. If one
    observes polytene chromosomes during
    the late prepupal stage, different bands
    appear to be puffed up. For 40 years, this
    has been understood to be the result of
    gene activity .

    Definition of polytene chromosome
    A giant chromosome produced by an endomitotic process
    in which, following synapsis of the two homologues,
    multiple rounds of replication produce chromatids that
    remain synapsed together in a haploid number of
    chromosomes. Polytene chromosomes have been studied
    mostly in Drosophila salivary glands, in which
    chromosomes undergo 10 cycles of replication without
    separation of the daughter chromosomes. This leads to
    1024 identical strands of chromatin aligned side by side.
    These chromosomes are easy to see with a light
    microscope because of their large size and precise
    alignment. The chromosomes are seen as distinct
    alternating dark and light bands which has given rise to a
    polytene chromosome map. The dark bands correspond to
    more condensed regions of the chromatin, and the light
    (interband) regions are less dense regions.

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