We used the Quant-iT™ RiboGreen® RNA Assay Kit (Life Technologies, catalogue no. R11490) to determine the RNA yields from each 1.0 mL of CSF (n = 2 duplicates/kit). We utilised the low-range (0–50 pg/µL) protocol adopted for 200 µL samples in a 96-well format (Costar). The 96-well plates assays were read on a plate reader at each site. OHSU used a Molecular Devices SpectraMax M2 Plate Reader with SoftMax Pro 6 Analysis Software; TGen used a BioTek Cytation 3 Imaging Reader BioTek with Gen5 Software v. 2.06 BioTek; and UCSD used a Tecan Infinite 200 Plate Reader with Tecan i-control software v. 1.10.4.0.
I control software v 1
Manufactured by Tecan
Sourced in Switzerland
I-control software v. 1.10.4.0 is a laboratory software application developed by Tecan. The software's core function is to provide users with the ability to control and operate various Tecan laboratory instruments and devices.
Automatically generated - may contain errors
2 protocols using i control software v 1
1
Quantification of RNA in CSF Samples
2
Measuring Growth and Aggregation of M. catarrhalis
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M. catarrhalis strains were grown overnight on BHI agar, then subcultured into BHI broth the following day and grown to mid-log phase. Cells were then diluted to OD600 = 0.05 (or 0.1 for CDM) and 0.5 mL aliquots dispensed into 48-well tissue culture plates. Plates were grown with aeration overnight at 37 °C in a Tecan Infinite 200 Pro plate reader (Tecan, Switzerland) and read at 15 minute intervals for growth curve measurements (Tecan i-control software v 1.10.4.0). Assays were carried out in at least triplicate biological repeats.
For aggregation assays, cells were grown with aeration in BHI broth, and standardised to OD600 = 1.5. 2 mL of these suspensions were transferred to sterilised 8 mL cuvettes (Sarstedt), then cuvettes were sealed and incubated under stationary conditions. OD600 measurements were taken at periodic intervals above the level of cell sedimentation.
For aggregation assays, cells were grown with aeration in BHI broth, and standardised to OD600 = 1.5. 2 mL of these suspensions were transferred to sterilised 8 mL cuvettes (Sarstedt), then cuvettes were sealed and incubated under stationary conditions. OD600 measurements were taken at periodic intervals above the level of cell sedimentation.
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