Bacterial Viability Assay with RNase

Bacterial viability assays were performed using a Live/Dead BacLight bacterial viability kit (BacLight; Molecular Probes) as previously described.^{6 (link),21} 10⁵ CFU/mL of stained bacteria were incubated with RNase 7, human RNase 6, or mouse RNase 6. Changes in fluorescent intensity were measured using the Spectramax M2 multimode microplate reader (Molecular Devices, Sunnyvale, CA).^{6 (link),21} These results were compared with a standard curve that was generated using increasing concentrations of live:dead bacteria.

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Becknell B., Eichler T., Beceiro S., Li B., Easterling R., Carpenter A.R., James C., McHugh K.M., Hains D.S., Partida-Sanchez S, & Spencer J.D. (2014). Ribonucleases 6 and 7 have antimicrobial function in the human and murine urinary tract. Kidney international, 87(1), 151-161.

Publication 2014

Live/Dead BacLight Spectramax M2

Assays Bacteria Bacterial viability Devices Human Molecular probes Mouse rnase 6 Rnase Rnase 7

Corresponding Organization : Nationwide Children's Hospital

Other organizations : The Ohio State University, Le Bonheur Children's Hospital

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Variable analysis

independent variables

RNase 7
Human RNase 6
Mouse RNase 6

dependent variables

Changes in fluorescent intensity

control variables

10^5 CFU/mL of stained bacteria

controls

Standard curve generated using increasing concentrations of live:dead bacteria

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