Nucleic acid chemistry

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Author:
khaengel
ID:
265962
Filename:
Nucleic acid chemistry
Updated:
2014-03-11 10:58:42
Tags:
MCDB chapter
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Chapter 8
Description:
MCDB chapter 8
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  1. Describe DNA denaturation?
    DNA denaturation (melting): 80oC and pH extremes, hydrogen bonds between bases are broken, base stacking is disrupted, DNA is unwound and strands separate. It can be partial or complete denaturation and there is only a change in conformation. Each species has a characteristic denaturation temperature. The higher the GC content, the higher the denaturation temperature (observed by electron microscopy).
  2. Describe renaturation:
    Renaturation (annealing): (A) if some small parts of the two DNA strands are still attached they anneal in one rapid step (B) if strands are completely separated (1) SLOW, strands randomly collide (2) FAST, strands zipper.
  3. What can you learn about nucleic acid structure using heating and annealing?
    What you can learn about nucleic acid structure using heating and annealing: Melt human and mouse DNA, some sequences will anneal over a few hours. Relative number of hybrid duplexes means more sequence homology. One would look for percent hybridization to quantify sequence homology between DNA species.
  4. What can you learn about nucleic acid structure using UV light?
    What you can learn about nucleic acid structure using ultraviolet light: UV light causes permanent changes in DNA. Thymine dimers form on the same strand; a C5 C6 link (kink) forms and distorts base stacking due to covalent bonding.
  5. What can you learn about nucleic acid structure using hybridization?
    What you can learn about nucleic acid structure using hybridization: show that a DNA sequence is present (via southern blot) or identify a gene sequence in a whole genome or plasmid (via DNA microarray).
  6. What are the specific damages that UV light can cause to DNA/RNA?
    • All nucleotides absorb light at wavelength
    • maxima at 260 nm.
    • UV light causes permanent changes in DNA.
    • Thymine dimers form on the same strand; a C5 C6 link (kink) forms and distorts
    • base stacking due to covalent bonding.

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