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mBio. 9, e02073-16. A bioengineered three-dimensional cell culture platform integrated with microfluidics to address antimicrobial resistance in tuberculosis 2017

Bielecka, M.K., Tezera, L.B., Zmijan, R., Drobniewski, F., Zhang, X., Jayasinghe, S., and Elkington, P. 

Notes: Human PBMC were isolated and encapsulated in a alginate-collagen matrix (via electrostatic bead generator) either alone or after infection with Mycobacterium tuberculosis. Viability of PBMC-alone or M. tuberculosis-infected PBMC microspheres were measured with the CellTiter-Glo® 3D Cell Viability Assay. Luminescence measurements in the study were accomplished with either the GloMax® 20/20 Luminometer or GloMax® Discover Instrument. (4815)

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J. Virol. 91(23), e01455–17. Genome-Wide Mutagenesis of Dengue Virus Reveals Plasticity of the NS1 Protein and Enables Generation of Infectious Tagged Reporter Viruses. 2017

Eyre, N.S., Johnson, S.M., Eltahla A.A., Aloi, M., Aloia, A.L., McDevitt, C.A., Bull, R.A. and Beard, M.R.

Notes: An insertional mutagenesis screen of the Dengue virus genome was used to identify regions tolerating small 15 amino acid insertions. The NS1 protein, which is essential for viral genome replication, was shown to be highly tolerant to insertions. NS1 was tagged with both a minimal fusion tag (Small BiT) and NanoLuc Luciferase. Tagged protein variants were assessed for viral infectivity, and used to investigate the protein localization and levels of NS1. (5075)

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BMC Genomics 16, 986. The effect of heterogeneous transcription start sites (TSS) on the translatome: implications for the mammalian cellular phenotype. 2015

Dieudonné, F.-X., O’Connor, P.B.F., Gubler-Jaquier, P., Yasrebi, H., Conne, B., Nikolaev, S., Antonarakis, S., Baranov, P.V. and Curran, J.

Notes: A bicistronic firefly/Renilla construct was engineered and transfected into 1.25 × 105 MCF7 cells/well of a 12-well plate using the ViaFect™ Transfection Reagent. Reporter activities were measured with the Dual-Luciferase® Reporter Assay System and read on a GloMax® 20/20 Luminometer. (4688)

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J. Cell Sci. 127, 5261-72. Epiprofin orchestrates epidermal keratinocyte proliferation and differentiation. 2014

Nakamura, T., Yoshitomi, Y., Sakai, K., Patel, V., Fukumoto, S. and Yamada, Y.

Notes: Epiprofin (Epfn) was expressed in HaCaT cells from various HaloTag® Fusion vectors with either full-length or deletion mutants of the CMV promoter to provide different levels of Epfn expression. (The authors do not state whether the N-terminal or C-terminal HaloTag® Fusion vectors were used). HaCaT 72hr proliferation was highest when using the CMVd3 promoter (e.g., pFC17A or pFN24A) for the lowest expression level. Reporter assays were performed on in the HaCaT cells as well and were measured with the Dual-Luciferase® Reporter Assay System on a GloMax® Instrument. Transfection studies performed with primary keratinocytes utilized the ViaFect™ Transfection Reagent using the reverse transfection method. (4690)

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Anal. Biochem. 423(2), 224-228. A bioluminescence assay for DNA methyltransferase activity based on methylation-resistant cleavage. 2012

Jiang, C., Yan, C.Y., Huang, C., Jiang, J.H., and Yu, R.Q.

Notes: This paper describes a bioluminescence-based method for the detection of DNA methyltransferase activity based on methylation-resistant cleavage and protein expression. The authors used Dam methylase as a model enzyme, MboI as the methylation-resistant endonuclease, and luciferase reporter DNA (LR-DNA) as the target. LR-DNA was amplified by PCR and then used as substrate in a Dam methylation reaction. If the target sites in the LR-DNA were fully methylated, they were resistant to subsequent MboI cleavage and expressed luciferase upon in vitro transcription/translation. Incomplete methylation or the absence of methylation resulted in DNA digestion and diminished/absent luciferase activity. TNT® T7 Quick for PCR DNA was used for in vitro translation of the LR-DNA, and the Luciferase Assay System and GloMax® 20/20 Luminometer were used to measure luciferase activity. (4191)

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Talanta 85, 527–32. Improvement of the analysis of the biochemical oxygen demand (BOD) of Mediterranean seawater by seeding control. 2011

Simon, F.X., Penru, Y., Guastalli, A.R., Lorens, J. and Baig, S.

Notes: Biochemical oxygen demand (BOD) is useful for determining water biodegradability, and the application of this measure allows researchers to analyze the efficiency of a given water treatment process in reducing the biodegradable natural organic matter (NOM). Such analyses allow researchers to determine the amount of oxygen required for the biochemical degradation of organic material over time. BOD protocols are well established for freshwater and wastewater; however, no satisfactory standard protocols have been developed for seawater and saltwater. The authors of this study set out to develop a protocol for a reliable Mediterranean seawater analysis using an appropriate seeding method. Raw seawater was collected from the Mediterranean Sea at the desalination plant at El Prat de Llobregat (Spain). The BacTiter-Glo™ Microbial Cell Viability Assay, which uses ATP indicator as the presence of viable cells, was used to monitor inoculum activity. Assay results were measured using the GloMax® 20/20 Luminometer. The authors determined a minimum seed quantity required for reliable BOD determination. (4260)

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J. Water Research 44, 3915-23. Measurement and interpretation of microbial adenosine triphosphate (ATP) in aquatic environments. 2010

Hammes, F., Goldschmidt, F., Vital, M., Wang, Y., and Egli, T.

Notes: In this study the luminescence-based BacTiter-Glo System and the GloMax 20/20 luminometer were used to detect ATP concentrations as low as 0.0001 nM in various water samples. The results correlated with those generated using traditional microbial detection methods. (4103)

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J. Microbiology 155, 1310-17. Solar disinfection (SODIS) and subsequent dark storage of Salmonella typhimurium and Shigella flexneri. 2009

Bosshard, F., Berney, M. and Scheifele, M.

Notes: In this study, the effect of solar disinfection on Shigella flexneri and Salmonella typhimurium in drinking water samples was evaluated. A variety of viability indicators were used to investigate the effectiveness of the disinfection method, including measurement of cellular ATP levels. The BacTiter-Glo Assay was used for ATP detection. (4104)

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