title: A novel approach for extraction of PCR-compatible DNA from activated sludge samples collected from different biological effluent treatment plants.
abstract: This paper describes a method that facilitates the extraction of PCR-compatible DNA from different activated sludge samples. The approach involves a novel preprocessing step in DNA extraction, which removes potential PCR inhibitors. The sludge was washed with different ratios of acetone and petroleum ether after pretreatment with 0.01% Tween-20 at 50 degrees C. It was observed that an initial washing step with 50 mM Tris-HCl, pH 9.0, before the detergent-solvent step, improved the quality of the extracted DNA. The extraction protocol resulted in amplifiable amounts of DNA when 10 mg of a sludge sample was used, even in the presence of phenol as a sludge contaminant. The usefulness of the extracted template was demonstrated by carrying out different PCR reactions. The random amplified polymorphic DNA (RAPD) patterns demonstrated the diversity of sludge samples.
authors.affiliation: National Environmental Engineering Research Institute, Nehru Marg, Nagpur 44 0020, India. hemantdrd@hotmail.com
authors.forename: Hemant J
authors.initials: HJ
authors.lastname: Purohit
authors.forename: Atya
authors.initials: A
authors.lastname: Kapley
authors.forename: Aditi A
authors.initials: AA
authors.lastname: Moharikar
authors.forename: Gurpreet
authors.initials: G
authors.lastname: Narde
mesh_terms: D004269:DNA, Bacterial / Q000032:analysis* / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007220:Industrial Waste; D016133:Polymerase Chain Reaction / Q000379:methods; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D012722:Sewage / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial; D007220:Industrial Waste; D012722:Sewage