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DNA protocols

DNA storage Clean and durty How to remove DNA?  Comments

links RNase treatment DNA shearing NA purification DNA purification from mammalian tissues DNA purification from cell culture

DNA storage

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





Clean and durty

clean durty genomic DNA, cDNA PCR-amplified material library without adapters library after adaptor ligation unopened tubes with PCR reaction 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.
• Typical PCR reaction.
• Amplification with urididne 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 "durty".
• Use separate pen, markers, etc. in durty area. Do not use laboratory journal in durty area.
• Remove/clean contamination-dangerous things as early as possible:
  • wash electrophoresis chambers and plates;
  • clean banch immediately after durty 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 tach anything by contaminated gloves/hands:
  • water source;
  • door handles;

How to remove DNA?

good choice bad choice buffer, which disolve DNA: whater, detergents 100% or 70% ethanol: DNA does not solubilize NH3-containing cleanoing solution, which strongly adsorb DNA



• hypochlorit Na, several minutes;