Orin C. Shanks, Catherine A. Kelty, Mano Sivaganesan, Manju Varma, and Richard A. Haugland Appl and Envir. Microbiol. 2009. Vol 75.

Overview

  • Key words: Waterborne disease, environmental waters, microbial community, DNA extraction
  • Aim of the study: Development of a method to assess microbial community present in waste water sample
  • Application: Quantitative PCR
  • Sample name: Wastewater
  • Material: FastPrep-24™ instrument, FastDNA™ Spin Kit for Soil containing Lysing Matrix E
  • Buffer: Sodium Phosphate Buffer and MT Buffer supplied with the FastDNA™ Spin Kit for Soil

Protocols & Parameters

500ml of primary effluent was collected and immediately stored on ice. 25 ml of each sample was filtered through a 0.2 -μm pore size supor-200 filters and each filter was placed in a sterile 1.5 ml microtube and stored at -80°C. For DNA extraction.


  1. Cut the freezed filters with a sterile cutter.
  2. Add the cutted filters to a Lysing Matrix E tubes.
  3. Add 978 μl of Sodium Phosphate Buffer and 122 μl of MT buffer, provided with the FastDNA™ spin kit for Soil.
  4. Homogenize in the FastPrep-24™ instrument for 120 seconds at a speed setting of 6.0
  5. Centrifuge at 14,000 x g for 5-10 minutes to pellet debris.
  6. Proceed with the FastDNA™ Spin Kit for Soil extraction protocol.

Conclusion

  • The FastPrep-24™ and associated matrices have demonstrated successful lysis and DNA extraction from 20 samples of wastewater in only 120 seconds.
  • This method saves hours of work during sample preparation, ensures high purified DNA and an effective PCR amplification.