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    Hyperchromic Effect

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    victor
    Single-stranded DNA absorbs more UV light than double-stranded DNA, known as the hyperchromic effect. When DNA is heated above its melting temperature, the double helix unwinds into single strands. This causes the bases to become unstacked and absorb more light, increasing absorbance by 37%. Hyperchromicity can be used to track DNA conditions as temperature changes. The melting temperature is where absorbance is halfway between single and double-stranded levels, indicating half the DNA is denatured.

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    Hyperchromic Effect

    Uploaded by

    victor
    2K views7 pages
    Single-stranded DNA absorbs more UV light than double-stranded DNA, known as the hyperchromic effect. When DNA is heated above its melting temperature, the double helix unwinds into single strands. This causes the bases to become unstacked and absorb more light, increasing absorbance by 37%. Hyperchromicity can be used to track DNA conditions as temperature changes. The melting temperature is where absorbance is halfway between single and double-stranded levels, indicating half the DNA is denatured.

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    Single-stranded DNA absorbs more UV light than double-stranded DNA, known as the hyperchromic effect. When DNA is heated above its melting temperature, the double helix unwinds into single strands. This causes the bases to become unstacked and absorb more light, increasing absorbance by 37%. Hyperchromicity can be used to track DNA conditions as temperature changes. The melting temperature is where absorbance is halfway between single and double-stranded levels, indicating half the DNA is denatured.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    2K views7 pages

    Hyperchromic Effect

    Uploaded by

    victor
    Single-stranded DNA absorbs more UV light than double-stranded DNA, known as the hyperchromic effect. When DNA is heated above its melting temperature, the double helix unwinds into single strands. This causes the bases to become unstacked and absorb more light, increasing absorbance by 37%. Hyperchromicity can be used to track DNA conditions as temperature changes. The melting temperature is where absorbance is halfway between single and double-stranded levels, indicating half the DNA is denatured.

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    © All Rights Reserved
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    Download as pptx, pdf, or txt
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    HYPERCHROMIC EFFECT

    BCH 323 PRESENTATION


    The absorption of single strand DNA is higher than the absorbance of double strand
    DNA , this is known as a hyperchromic effect (means: more colour).
    Hyperchromicity is the increase of absorbance (optical density) of a material.
    The most famous example is the hyperchromicity of DNA that occurs when the DNA
    duplex is denatured.
    The UV absorption is increased when the two single DNA strands are being separated,
    either by heat or by addition of denaturant or by increasing the pH level.
    The opposite, a decrease of absorbance is called hypochromicity.
    Heat denaturation of DNA, also called melting, causes the double helix structure to
    unwind to form single stranded DNA. When DNA in solution is heated above its melting
    temperature (usually more than 80 C), the double-stranded DNA unwinds to form
    single-stranded DNA.
    The bases become unstacked and can thus absorb more light. In their native state, the
    bases of DNA absorb light in the 260-nm wavelength region. When the bases become
    unstacked, the wavelength of maximum absorbance does not change, but the amount
    absorbed increases by 37%. A double strand DNA dissociating to single strands produces
    a sharp cooperative transition.
    Hyperchromicity can be used to track the condition of DNA as temperature changes. The
    transition/melting temperature (T) is the temperature where the absorbance of UV light
    is 50% between the maximum and minimum, i.e. where 50% of the DNA is denatured.
    The hydrogen bonds between the paired bases in the double helix limits the resonance
    behavior of the aromatic ring of the bases which results in decrease in the UV
    absorbance of double stranded DNA (hypochromic effect), while in singe stranded DNA
    the bases are in free form and don't form hydrogen bonds with complementary bases
    which results in 40% higher absorbance in single stranded DNA (hyperchromic) at the
    same concentration.
    The Hyperchromic effect is the striking increase in absorbance of DNA upon
    denaturation. The two strands of DNA are bound together mainly by the stacking
    interactions, hydrogen bonds and hydrophobic effect between the complementary
    bases.
    Some important concepts in Hyperchromic Effect

    At Tm, the DNA is half denatured and half double stranded.


    By lowering the temperature below the Tm, the denatured DNA strands would anneal back
    into a double stranded DNA.
    When temperature is above the Tm, the DNA is denatured
    Because the melting temperature (Tm), occurs almost instantly at a certain T, monitoring
    the absorbance of the DNA at various temperature would indicate the melting T.
    By being able to find the temperature at which DNA melted and annealed, scientists are
    able to separate DNA strands and anneal them with other DNA strands. This is important in
    creating hybrid DNAs, which consists of two DNA strands from different sources. Since DNA
    strands can only anneal if they are similar, the creation of hybrid DNAs can indicate
    similarities between genomes of different organisms.
    THANK YOU!

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