DNA

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[править] DNA storage

  • normal storage: -20°C
  • very long (>200kb) DNA should not be freezed. Store it at +4°C. NB! microbial degradation may happens at this temperature
  • salt/ethanol precipitate is a safe way of transportation at ambient temperature
  • DNA is chemically unstable in acidic solutions




[править] Contamination-dangerous DNA

cleandirty
  • genomic DNA, cDNA
  • library without adapters
  • unopened tubes with PCR reaction
  • PCR-amplified material
  • library after adaptor ligation
  • gel with fractionated adapter-ligated or preamplified library


  • "PCR-safe" and "sterile" work significantly differ from each other.
  • Use reasonable aliquotes of reagents. If contamination happens, through them away.
  • Amplification with dUTP and UDGase digestion does not prevent contamination completely. It decrease contamination level on ~3 orders of magnitude.
  • Try to organize a day work from "clean" to "dirty".
  • Use separate pen, markers, etc. in dirty area. Do not use laboratory journal in dirty area.
  • Remove or clean contamination-dangerous things as early as possible:
    • wash electrophoresis chambers and plates;
    • clean bench immediately after dirty work (gel-band cutting, ePCR transfer, etc.);
    • wash hands and gloves.
  • Spin down (for at least 5 sec) all contamination-dangerous tubes before opening.
  • Do not touch anything by contaminated gloves/hands:
    • water source;
    • door handles.



[править] How to remove DNA?

Bad choice: 100% or 70% ethanol. Because DNA does not solubilize in it.

Good choice:

  • whater, may be with detergents. DNA is easily disolvable in this buffer.
  • NH3-containing cleaning solution. After washing by such solution surfaces strongly (irreversibly) adsorb DNA.
  • hypochlorit Na, several minutes. Chemical degradation of DNA.




Источник — «http://molbiol.ru/wiki/DNA»


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