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    Dna Denaturation and Renaturation

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    Soumyaranjan Pati
    Denaturation of DNA involves breaking the hydrogen bonds between the two strands, leaving the covalent bonds intact. This separates the double helix into single strands. Denaturation can be caused by heat (over 90°C), sodium hydroxide, or high salt concentrations. Effects include increased UV absorption and decreased viscosity as the double helix unwinds. Renaturation occurs when conditions return to normal, allowing the single strands to rewind into the original double helix structure.

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    Dna Denaturation and Renaturation

    Uploaded by

    Soumyaranjan Pati
    131 views2 pages
    Denaturation of DNA involves breaking the hydrogen bonds between the two strands, leaving the covalent bonds intact. This separates the double helix into single strands. Denaturation can be caused by heat (over 90°C), sodium hydroxide, or high salt concentrations. Effects include increased UV absorption and decreased viscosity as the double helix unwinds. Renaturation occurs when conditions return to normal, allowing the single strands to rewind into the original double helix structure.

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    Denaturation of DNA involves breaking the hydrogen bonds between the two strands, leaving the covalent bonds intact. This separates the double helix into single strands. Denaturation can be caused by heat (over 90°C), sodium hydroxide, or high salt concentrations. Effects include increased UV absorption and decreased viscosity as the double helix unwinds. Renaturation occurs when conditions return to normal, allowing the single strands to rewind into the original double helix structure.

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

    Dna Denaturation and Renaturation

    Uploaded by

    Soumyaranjan Pati
    Denaturation of DNA involves breaking the hydrogen bonds between the two strands, leaving the covalent bonds intact. This separates the double helix into single strands. Denaturation can be caused by heat (over 90°C), sodium hydroxide, or high salt concentrations. Effects include increased UV absorption and decreased viscosity as the double helix unwinds. Renaturation occurs when conditions return to normal, allowing the single strands to rewind into the original double helix structure.

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    Denaturation of DNA:In the denaturation process, the hydrogen bonds between

    two strands are broken giving rise to two single strands. The covalent bonds of DNA remain
    unaffected.

    Denaturation of DNA double helix takes place by the


    following denaturating agents:

    (i) Denaturation by Temperature:

    If a DNA solution is heated to approximately 90°C or above there will


    be enough kinetic energy to denature the DNA completely causing it to
    separate into single strands. This denaturation is very abrupt and is
    accelerated by chemical reagents like urea and formamide.The
    chemicals enhance the aqueous solubility of the purine and pyrimidine
    groups. This separation of double helix is called melting as it occurs
    abruptly at a certain characteristic temperature called denaturation
    temperature or melting temperature (Tm).It is defined as temperature
    at which 50% of the DNA is melted.If several samples of DNA are
    melted, it is found that the Tm is highest for those DNAs that contain
    the highest proportion of G—C. Actually the value is used to estimate
    the percentage of G—C in a DNA sample. In fact, the Tm of DNA from
    many species varies linearly with G—C content.

    This relationship between Tm and G—C content arises due to guanine


    and cytosine form three hydrogen bonds when base paired, whereas
    adenine and thymine form only two.

    DNA Denaturation through NaOH Treatment


    Apart from heating, chemical denaturation can also be achieved through the use of NaOH. A certain
    concentration of NaOH can be used to denature DNA safely, often in as little as one minute. As the
    concentration of NaOH used is reduced, the process will take longer -- but the DNA can still be fully
    denatured.

    DNA Denaturation through Salt .


    A high concentration of salt will cause DNA to naturally denature, given the right concentration of
    salt. DNA denaturation with salts are similar to denaturation through the use of organic solvents. In
    general, DNA denaturation through salt cannot be renatured.

    Effect of denaturation of DNA:

    ● Increased absorption of UV light at 260nm wavelengths. The rate of absorption is directly


    proportional to the rate of denaturation
    ● Viscosity decreases, which reflects the physical change occurred in the DNA structure

    What is Renaturation?
    Renaturation is also known as annealing. When the temperature and pH return to optimum biological
    level, the unwound strand of DNA rewind and give back the dsDNA.

    If the DNA is not completely denatured, the renaturation process is fast and a one-step process, but
    if the DNAs are completely denatured then the renaturation process occurs in a two-step process.
    First complementary strands come together by random collision and then rewinding takes place
    forming a double helix.

    Renaturation occurs when the denatured DNAs are cooled in suitable conditions. Renaturation also
    depends on temperature, pH, length and constituents of the DNA structure. The renaturation rate is
    directly proportional to the number of complementary sequences present.

    With renaturation, absorption of UV (260nm) decreases and viscosity increases again.

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