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    ASAM NUKLEAT SFB (Pertemuan 12) Coretan

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    Ajenglestari Wicaksono
    The document discusses the structure and function of biomolecules, focusing on nucleic acids. It describes the components of nucleic acids including pentose sugars, nitrogen bases, and phosphate groups. It also explains the structures of RNA, DNA, and the double helix structure of DNA.

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    ASAM NUKLEAT SFB (Pertemuan 12) Coretan

    Uploaded by

    Ajenglestari Wicaksono
    16 views51 pages
    The document discusses the structure and function of biomolecules, focusing on nucleic acids. It describes the components of nucleic acids including pentose sugars, nitrogen bases, and phosphate groups. It also explains the structures of RNA, DNA, and the double helix structure of DNA.

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    ASAM NUKLEAT SFB (Pertemuan 12) coretan

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    The document discusses the structure and function of biomolecules, focusing on nucleic acids. It describes the components of nucleic acids including pentose sugars, nitrogen bases, and phosphate groups. It also explains the structures of RNA, DNA, and the double helix structure of DNA.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    16 views51 pages

    ASAM NUKLEAT SFB (Pertemuan 12) Coretan

    Uploaded by

    Ajenglestari Wicaksono
    The document discusses the structure and function of biomolecules, focusing on nucleic acids. It describes the components of nucleic acids including pentose sugars, nitrogen bases, and phosphate groups. It also explains the structures of RNA, DNA, and the double helix structure of DNA.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
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    You are on page 1of 51

    KI-2241

    Struktur dan Fungsi Biomolekul

    ASAM NUKLEAT
    Aditya Ayuwulanda, S.Pd., M.Si.
    aditya.ayuwulanda@ki.itera.ac.id
    Sistem informasi
    Meliputi: penyimpanan,
    penerimaan, pemrosesan,
    dan penyampaian
    informasi biologi
    Asam nukleat
    Komponen Asam Nukleat

    RNA DNA
    Komponen Asam Nukleat
    Gula Pentosa
    Komponen Asam Nukleat
    BASA NITROGEN
    RA RB
    Rantai DNA
    Pembentukan Ikatan Pospodiester pada
    Polinukleotida
    RNA

    DNA
    Heliks rangkap DNA
    • Dua rantai polinuklotida digulung sekitar
    sumbu
    • Tulang punggung gula-fosfat di luar, basa
    purin dan primidin di dalam
    • Basa hampir tegak lurus ke sumbu
    pengulangan heliks 34 Å/10 basa
    • Diameter 20 Å
    Watson-Crick model for structure of DNA
    Bentuk Rantai helix DNA
    STRUKTUR SEKUNDER ASAM NUKLEAT

    Struktur sekunder
    terjadi karena adanya
    interaksi hydrogen
    antara basa nitrogen
    pada asam nulekat
    Palindromic DNA (or RNA)
    STRUKTUR TERSIER ASAM NUKLEAT

    Circular DNA
    TEKNIK ISOLASI ASAM NUKLEAT
    Identifikasi DNA
    Agarose Gel Electrophoresis
    • Elektroforesis merupakan cara untuk memisahkan fraksi campuran
    berdasarkan pergerakan partikel bermuatan di bawah pengaruh
    medan listrik.

    • Elektroforesis umumnya digunakan untuk menentukan berat molekul


    protein maupun asam nukleat
    • DNA is negatively charged.
    • When placed in an electrical field, DNA will migrate toward the positive
    pole (anode).
    • An agarose gel is used to slow the movement of DNA and separate by
    size.
    H O2
     

    DNA

    - +

    Power • Polymerized agarose is porous,


    allowing for the movement of DNA
    Scanning Electron Micrograph
    of Agarose Gel (1×1 µm) 
    How fast will the DNA migrate?
    strength of the electrical field, buffer, density of agarose gel…
    Size of the DNA!
    *Small DNA move faster than large DNA
    …gel electrophoresis separates DNA according to size

    DNA

    small
    large
    - +

    Power
    Within an agarose gel, linear DNA migrate inversely
    proportional to the log10 of their molecular weight.
    Place the gel in the electrophoresis chamber.
    DNA

    buffer     
    wells
    Anode
    Cathode (positive)
    (negative)

    Add enough electrophoresis buffer to cover the gel to a depth of


    at least 1 mm. Make sure each well is filled with buffer.
    Sample Preparation

    Mix the samples of DNA with the 6X sample loading buffer (w/ tracking
    dye). This allows the samples to be seen when loading onto the gel, and
    increases the density of the samples, causing them to sink into the gel
    wells.

    6X Loading Buffer: 
     Bromophenol Blue (for color)
     Glycerol (for weight)
    Loading the Gel

    Carefully place the pipette tip over a well and gently expel the sample.
    The sample should sink into the well. Be careful not to puncture the
    gel with the pipette tip.
    Running the Gel

    Place the cover on the electrophoresis chamber, connecting the electrical


    leads. Connect the electrical leads to the power supply. Be sure the leads
    are attached correctly - DNA migrates toward the anode (red). When the
    power is turned on, bubbles should form on the electrodes in the
    electrophoresis chamber.
    Cathode
    (-)

     wells
    DNA  Bromophenol Blue
    (-)

    Gel

    Anode
    (+)

    After the current is applied, make sure the Gel is running in the correct
    direction. Bromophenol blue will run in the same direction as the DNA.
    Penentuan ukuran DNA
    Proses Denaturasi DNA
    • Proses denaturasi dapat dilakukan dengan
    pemanasan atau penambahan alkali
    • Proses denaturasi dapat diamati melalui pengukuran
    absorbansi pada spektrometri UV
    Renaturasi DNA
    • Proses renaturasi dapat terjadi jika konsentrasi garam
    (NaCl) dalam suspensi DNA cukup tinggi
    • Semakin besar konsentrasi DNA makan laju renaturasi
    akan semakin bertambah (karena probabilitas
    terjadinya tumbukan antar untai DNA semakin besar)

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