MilliporeSigma
  • Home
  • Search Results
  • Development of an Escherichia coli K12-specific quantitative polymerase chain reaction assay and DNA isolation suited to biofilms associated with iron drinking water pipe corrosion products.

Development of an Escherichia coli K12-specific quantitative polymerase chain reaction assay and DNA isolation suited to biofilms associated with iron drinking water pipe corrosion products.

Journal of water and health (2014-12-05)
Jingrang Lu, Tammie L Gerke, Helen Y Buse, Nicholas J Ashbolt
ABSTRACT

A quantitative polymerase chain reaction assay (115 bp amplicon) specific to Escherichia coli K12 with an ABI(TM) internal control was developed based on sequence data encoding the rfb gene cluster. Assay specificity was evaluated using three E. coli K12 strains (ATCC W3110, MG1655 & DH1), 24 non-K12 E. coli and 23 bacterial genera. The biofilm detection limit was 10(3) colony-forming units (CFU) E. coli K12 mL(-1), but required a modified protocol, which included a bio-blocker Pseudomonas aeruginosa with ethylenediaminetetraacetic acid buffered to pH 5 prior to cell lysis/DNA extraction. The novel protocol yielded the same sensitivity for drinking water biofilms associated with Fe3O4 (magnetite)-coated SiO2 (quartz) grains and biofilm-surface iron corrosion products from a drinking water distribution system. The novel DNA extraction protocol and specific E. coli K12 assay are sensitive and robust enough for detection and quantification within iron drinking water pipe biofilms, and are particularly well suited for studying enteric bacterial interactions within biofilms.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Iron, chips, 99.98% trace metals basis
Sigma-Aldrich
Iron, foil, thickness 0.1 mm, ≥99.9% trace metals basis
Sigma-Aldrich
Iron, wire, diam. 0.5 mm, ≥99.9% trace metals basis
Sigma-Aldrich
Iron, powder, −325 mesh, 97%
Sigma-Aldrich
Iron, rod, diam. 6.3 mm, 99.98% trace metals basis
Sigma-Aldrich
Iron, puriss. p.a., carbonyl-Iron powder, low in magnesium and manganese compounds, ≥99.5% (RT)
Iron, IRMM®, certified reference material, 0.5 mm wire
Iron, foil, light tested, 25x25mm, thickness 0.015mm, 99.85%
Iron, foil, not light tested, 100x100mm, thickness 0.006mm, 99.85%
Iron, foil, not light tested, 50x50mm, thickness 0.001mm, 99.85%
Iron, foil, not light tested, 50x50mm, thickness 0.0005mm, temporary acrylic support, 99.85%
Iron, foil, not light tested, 150x150mm, thickness 0.008mm, 99.85%
Iron, wire reel, 0.5m, diameter 0.13mm, hard, 99.99+%
Iron, wire reel, 2m, diameter 0.25mm, annealed, 99.99+%
Iron, wire reel, 10m, diameter 0.25mm, hard, 99.99+%
Iron, wire reel, 5m, diameter 0.25mm, annealed, 99.99+%
Iron, wire reel, 2000m, diameter 0.125mm, hard, 99.5%
Iron, wire reel, 0.5m, diameter 1.0mm, as drawn, 99.99+%
Iron, wire reel, 0.1m, diameter 0.5mm, hard, 99.99+%
Iron, wire reel, 0.05m, diameter 0.5mm, as drawn, 99.998%
Iron, rod, 200mm, diameter 50mm, as drawn, 98+%
Iron, tube, 200mm, outside diameter 10.6mm, inside diameter 10.0mm, wall thickness 0.3mm, as drawn, 99.8+%
Iron, foil, 10m coil, thickness 0.038mm, hard, 99.5%
Iron, rod, 500mm, diameter 9.5mm, as drawn, 98+%
Iron, foil, 300x300mm, thickness 0.25mm, hard, 99.5%
Iron, foil, 10m coil, thickness 0.5mm, hard, 99.5%
Iron, rod, 100mm, diameter 30mm, as drawn, 98+%
Iron, tube, 200mm, outside diameter 25.0mm, inside diameter 21mm, wall thickness 2mm, annealed, 99.5%
Iron, tube, 1000mm, outside diameter 8.0mm, inside diameter 5mm, wall thickness 1.5mm, annealed, 99.5%
Iron, rod, 500mm, diameter 12.7mm, as drawn, 99.95%