Protocol for decontamination of instruments, reagents and laboratory areas for challenging DNA analysis.

Summary or abstract

The protocol constitute a guideline for decontamination of laboratory and basic tools used during the work with the biological material for DNA analysis. It is published as a supplementary material to the December-February Special Issue on Biobanking in Emerging countries in the journal Biopreservation & Biobanking.

Full protective clothing (apron, mask, cap, gloves and protective shoes) is required at each stage of decontamination process. The protective clothing used during decontamination should be discarded immediately after the procedure! An optimal clean room facility is equipped with positive air-pressure, UV-light, HEPA-filtered air and LAF-benches.

DECONTAMINATION OF LABORATORY EQUIPMENT

Material: 15% bleach, 70% ethanol, dust-free paper

  1. Before entering the lab put on full protective clothing. Always put on the gloves first and change them frequently.
  2. Thoroughly clean all equipment in the laboratory (e.g. shakers, centrifuges) using 15% bleach and dust-free sheets of paper.
  3. After drying, clean the same equipment using 70% ethanol. Let the equipment dry.
  4. If necessary, repeat the cleaning procedure from step 2.
  5. Switch on the UV-light for at least 20 min. If the laboratory will be used for the first time after a long break or moving, apply UV light overnight.

DECONTAMINATION OF INSTRUMENTS, RACKS, MORTARS etc.

Material: container with a lid containing 15% bleach, lockable containers containing dH2O (x2), container with a lid containing dust-free sheets of paper.

  1. Place all of the instruments into the container with 15% bleach for 15-60 minutes.
  2. Place cleaned instruments into the first container with dH2O for at least 2 h.
  3. Place all the instruments into the second container with dH2O for at least 2 h.
  4. Transfer all the instruments into container with dust-free paper. Leave the instruments until they are completely dry.

Important! Please remember that metal instruments under the influence of chlorine will corrode and accumulate rust. In this case, the soaking time in bleach can be reduced depending on the resistance and material of a particular instrument.

SURFACE DECONTAMINATION OF BONES

  1. Only a small part of large bone are used and decontaminated. Therefore, the remainder of the bone must be wrapped in aluminum foil or elastic plastic. This will allow appropriate decontamination of the part designated for extraction and protect against the penetration of contamination from other parts of the bone.
  2. Remove the top layer of 2-3 mm of bone using sandpaper.
  3. Clean off the dust and powder using dH2O
  4. Clean the bone surface with 15% bleach.
  5. Clean the surface with 70% ethanol. 
  6. Clean the surface with dH2O. Let the bone dry. This part of decontaminated bone is ready for cutting the peace of sample to be used for DNA extraction.

Important! Change the gloves between each step of the decontamination process.

Citation

Buś M.M., Allen M. (2014). Collecting and preserving biological samples from challenging environments for DNA analysis. Biopreservation and Biobanking, 12:17-22

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DNA extraction and storage