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This assay was completed in 2 days.
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The homohopane index decreased in the first 60 days and increased from then on until the assay was completed (180 days), with an unusual homohopane biodegradation preference C33 > C32 > C31 > C34 > C35 Munoz et al. (1997) showed 25% depletion of C35 pentakishomohopane and only 3% depletion of C31 homohopanes in soils exposed to an oil spill for eight years.
Following incubation, the assay was completed as for the standard RIP.
The plates were incubated for 18 to 20 hours at 37°C in a humidified incubator with 5% CO2, and the assay was completed according to the manufacturer's directions.
After the assay was completed, the data were pooled for statistical analysis.
Foci larger than 2 μm were counted for each group (n=30) and measurements on inclusions were performed using NIH ImageJ™ software and the assay was completed at least 3 times similarly as performed in [ 102].
After the assay was completed, viable cells in each well were counted using a Vi-Cell XR cell counter and the cell counts used to normalize the OCR rates, with OCR being expressed as pmoles oxygen/minute/10 cells.
After peroxidase quenching with hydrogen peroxide (3 %) for 5 min, fixed cells were incubated with mouse anti-human CD68 (Abcam, Cambridge, MA) for 60 min, and the assay was completed with Dako's immunoperoxidase kit (Dako Corp., Carpintera, CA).
At this point changes to the assay were completed and validation was performed for the sequencing and large rearrangement components for the entire 25-gene panel test.
The hybridization assay was completed in 50 min.
The assay was completed within 2 h from DNA extraction.
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