Jörg D. Ettenauer, Guadalupe Piñar, Ksenija Lopandic, Bernhard Spangl, Günther
Ellersdorfer, Christian Voitl, Katja Sterflinger
Science of the Total Environment. 2012. Vol 439.

Overview


Protocols & Parameters

  1. Sampling was done using a sterile scalpel or an ethanol flamed hammer and a chisel to remove the material from the walls and collected it in sterile plastic bags.
  2. The transport and storage of the sampling material were done at room temperature.
  3. In the laboratory, samples were ground for 2 min in liquid nitrogen using a sterile mortar and pestle, collected in a sterile 50 ml falcon tube and homogenized by manual shaking.
  4. Three different sample amounts of each material, 50 mg, 100 mg and 250 mg (each in triplicate), were weighed in a Sartorius precision scale for each extraction method.
  5. Tubes were either immediately processed or stored at − 20 °C. The resulting nine samples for each method were further subjected to the different DNA extraction methods.
  6. When using the FastDNA™ Spin Kit for Soil method, samples were processed two times in the FastPrep® homogenizer for 40 s at a speed of 6 m/s.

Results

Representative examples of bacteria and fungal fingerprints obtained from the plaster material. The banding patterns of the three tested sample amounts (50,100,250 mg) from the FastDNA™ Spin kit for soil.


Conclusion

  • Up to thirteen extraction methods were evaluated with three building materials. The FastDNA™ Spin kit for soil showed to be the best DNA extraction method and could provide positive results for all tests with all tested samples.
  • This study shows that the FastPrep® extraction method is a gold standard for quantification of indoor fungi and bacteria in building materials.