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For immunoblots, 5µl aliquots from a total extraction volume of 12µl were spotted and fixed to a Nylon Hybond+ membrane.
Furthermore, ten participants had accumulated a total extraction volume of >100, among them four with an extraction volume of >400.
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Irrespective of exposure (total puff number) or nicotine extraction volume, all total deposited mass and nicotine data were normalised to surface area per puff.
To mass standardize the total chlorophyll concentrations, this data was multiplied by the extraction volume (1 ml) and divided by the fresh weight of the samples (mg chl/g FW).
With 2,380 bf representing half the volume of commercial timber on a typical hectare of suitable forest in Tambopata, the mean extraction intensity in each ecotourism concession was calculated at 381,251±518,386 bf yr−1 for a total extraction of 1,906,256 bf yr−1, equivalent to only 0.03% of the 2004 extraction intensity in Madre de Dios as a whole [40].
The RisN method [29] was preformed with doubled extraction volume.
Manual extraction was performed to obtain a final extraction volume of 200 µl DNA.
The extraction volume of HCl was adjusted proportionately.
Multiple plant and tissue samples were used for each extraction in a 12 ml extraction volume.
The assay mixture, consisting of 25 mM Orn, 25 mM α-ketoglutarate and 100 μl plant extract in a total volume of 500 μl extraction buffer, was incubated at 37°C for 20 min. The reaction was terminated by the addition of 150 μl 3 M HClO4.
In the case of swabs, reactions were performed on 100 ng of extracted DNA amplifying the bacterial 16S rRNA gene, and the results were normalised to the total volume of extraction buffer.
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