4609MA In the Literature Protocol overview of the BacTiter-Glo™ Microbial Cell Viability Assay. Sibag, M. et al. (2015) Interference sources in ATP bioluminescence assay of silica nanoparticle toxicity to activated sludge. J. Microbiol. Methods 113, 65–71. BacTiter-Glo™ Substrate Mix BacTiter-Glo™ Buffer BacTiter-Glo™ Reagent In this article, authors wanted to determine if the silica nanoparticles (SNP) interfered with microbial activity in activated sludge by measuring ATP as a proxy for bacterial viability. To test this hypothesis, samples of deionized (DI) water (no-ATP control), SNP suspension (100mg/l), fresh activated sludge and autoclaved activated sludge were mixed with an equal volume of the BacTiter-Glo™ Microbial Cell Viability Assay Reagent, and luminescence measured using a GloMax® 20/20 Luminometer. Further testing was performed to assess if SNPs alone, cell debris or synthetic sewage would interfere with the ATP assay by comparing ATP standard prepared in water with SNPs, autoclaved activated sludge or synthetic sewage with ATP standard. Sample pretreatment (e.g., centrifigation of sample) prior to ATP measurement was tested to see if assay interference could be reduced. Justo, A. et al. (2015) BAC filtration to mitigate micropollutants and EfOM content in reclamation reverse osmosis brines. Chemical Engineering Journal 279, 589–96. Mixer Luminometer The authors were interested in how effective biologically activated carbon (BAC) filters, which are granular activated carbon (GAC) filters colonized by microbial biomass, were in removing unwanted pollutants from reverse osmosis (RO) discharge during waste water treatment. To measure the amount of ATP, a proxy for biological activity, BacTiter-Glo™ Microbial Cell Viability Assay Reagent was added to 100µl of sample, mixed for 3 minutes and luminescence measured using a GloMax® 20/20 Luminometer. The amount of ATP was measured before and after sample filtration through a 0.22μm filter to quantify total and free ATP respectively. To estimate the amount of ATP contained within the biofilms growing on the BAC filters, 200mg of GAC and 100μl of phosphate buffer were added to a tube, mixed for 3 minutes in an orbital shaker before adding 300μl of BacTiter-Glo™ Reagent. After mixing for another 3 minutes, 200μl of the reaction was transferred to a new tube and luminescence was measured. Ordering Info More BacTiter-Glo™ Citations » enews | August 2015 Pagina 8
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