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Southern Blotting exam
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  • Southern Blotting
    A technique used to detect specific DNA sequences using radioactive probes.
  • Radioactive Probes
    Single-stranded DNA molecules that are radioactively labeled and bind to complementary sequences.
  • What is the purpose of gel electrophoresis in Southern blotting?
    To separate DNA fragments by size.
  • Denaturation
    The process of converting double-stranded DNA into single-stranded DNA.
  • What is the role of the nitrocellulose filter paper in Southern blotting?
    To transfer and immobilize single-stranded DNA from the gel.
  • How are DNA fragments visualized in Southern blotting?
    By using radioactive probes that bind to specific sequences, visualized as bands on the filter paper.
  • Incubation with Radioactive Probes
    The step where the nitrocellulose filter paper is exposed to DNA probes to detect specific sequences.
  • What does the presence of bands on the filter paper indicate?
    The presence of specific DNA sequences of interest.
  • Gel Electrophoresis
    A technique used to separate DNA fragments based on size.
  • What is the first step in Southern blotting?
    Fragmenting unknown DNA samples and separating them via gel electrophoresis.
  • Denaturing Buffer
    A solution used to denature DNA, making it single-stranded.
  • What happens during the denaturation step in Southern blotting?
    DNA is converted to single-stranded DNA by increasing the pH.
  • Blotting
    The process of transferring DNA from the gel to the nitrocellulose filter paper.
  • What is the final step in Southern blotting?
    Analyzing the radioactive filter paper to visualize the DNA bands.
  • Complementary Sequences
    DNA sequences that can pair with each other via base pairing.
  • What is the significance of the yellow color in the radioactive probe example?
    It represents the presence of radioactivity, indicating the sequence of interest.
  • Northern Blotting
    A technique used to detect RNA sequences.
  • Western Blotting
    A technique used to detect specific proteins.
  • What is the role of paper towels in the denaturation step?
    To absorb the denaturing buffer as it migrates upwards.
  • What does the term 'blotting' refer to in Southern blotting?
    Transferring DNA from the gel to the filter paper.
  • Single-Stranded DNA (ssDNA)
    DNA that has been denatured to separate its two strands.
  • What is the purpose of using a sponge in the denaturation step?
    To help absorb the denaturing buffer and facilitate its migration.
  • What does the presence of radioactivity in a sample indicate?
    That the sample contains the specific DNA sequence of interest.
  • What is the main use of Southern blotting?
    To detect specific DNA sequences for genetic analysis and research.
  • What happens to the DNA in the gel after denaturation?
    It is transferred to the nitrocellulose filter paper.
  • What is the role of the denaturing buffer in Southern blotting?
    To denature DNA by increasing the pH, making it single-stranded.
  • What is visualized as bands on the filter paper in Southern blotting?
    The DNA sequences that are complementary to the radioactive probes.
  • What is the significance of lanes 2 and 4 in the final step of Southern blotting?
    They contain the DNA sequences of interest that are complementary to the probe.