Biochem Molecular Analyses (5)

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pctran90
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223994
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Biochem Molecular Analyses (5)
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2013-06-15 23:15:42
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Biochem Molecular Analyses
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Biochem Molecular Analyses
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  1. Antibody
    A protein produced by the vertebrate immune system that binds specifically and tightly to a small segment of a single protein.
  2. Antigen
    The protein recognized by an antibody.
  3. cDNA
    DNA complementary to all the mRNA present in the sample and DNA complementary to all the mRNA present in normal tissue used as probes in Microarray Analysis.
  4. Cloning
    The isolation of genomic fragments and replication of the isolated fragments into many copies.
  5. Denature
    The separation of DNA strands. (ie. due to changes in pH)
  6. Dideoxynucleotide Triphosphates
    Nucleotide triphosphates incorporated at the end of a growing DNA chain that causes chain termination because they lack the 3'-OH group. Used in DNA sequence determination.
  7. Gel Electrophoresis
    • An electric mechanism for separating DNA/RNA molecules based on their size. 
    • *DNA is negatively charged, so it moves towards the positive end.
    • *Smaller DNA fragments will move more quickly than larger molecules.
  8. Hybridization
    Refers to the base paring of the probe with any DNA fragment having a complementary sequence.
  9. Western Blot
    Protein blotting with an antibody probe that binds to specific target proteins.
  10. Immunoblotting
    The appropriate term for Western blotting/protein blotting.
  11. Karyotype
    The direct visualization of the complete set of chromosomes. Used to detect large changes in chromosome structure and number.
  12. Microarray
    • A technique to simultaneously assess the relative level of expression of all transcribed genes in a cell.
    • (Hint: Involves cDNA probes)
  13. Northern Blot
    A technique that analyzes mRNA and is used to examine the regulation of expression of specific genes. (Also used to examine the size of mRNA.)
  14. Oligonucleotide
    A short, single-stranded nucleic acid polymer.
  15. Plasmid
    Small, circular DNA molecule found in bacteria that exists separate from the bacterial genome.
  16. Vector
    An agent, such as a plasmid or a virus, that can incorporate foreign DNA and transfer that DNA from one organism to another.
  17. Polymerase Chain Reaction (PCR)
    A highly specific, sensitive, and widely applicable technique for detecting and quantitating DNA/RNA involving heating and cooling cycles.
  18. Probe
    A radioactive DNA molecule complementary to the specific DNA fragment of interest.
  19. Restriction Endonuclease
    Bacterial enzymes that cleave DNA at specific sequences.
  20. Recognition Sequence
    Specific 4-8 bp palindromic DNA sequences recognized by Restriction Endonucleases.
  21. Secondary Antibody
    An antibody that specifically binds to a primary antibody allowing visualization of bound antibodies of interest in immunoblotting.
  22. Sodium Dodecyl Sulfate (SDS)
    A detergent used to cause proteins to migrate through gels at rates proportional to only their size. It works by denaturing the proteins and coating them with a uniform negative charge.
  23. Southern Blot
    A technique that analyzes genomic DNA and detects large deletions, insertions, and sequence rearrangements.
  24. Synthetic Oligonucleotide Primers
    Short, synthetic single-stranded nucleic acid polymers used primers for DNA Polymerase.
  25. FISH
    A karyotyping technique in which a fluorescent dye that hybridizes to a specific chromosomal site is coupled a DNA molecule.
  26. "Staggered" Cleavage
    A mechanism in which dsDNA is cleaved that creates an overhang (sticky end).
  27. "Blunt" Cleavage
    A mechanism in which dsDNA is cleaved precisely in the center of the recognition sequence, leaving a flush end.

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