The calculation provided in the protocol accounts for sample background by subtracting the sample background blank.
The working reagent should be made fresh and used within 15 minutes.
If your samples have very low ODs you may be below the kit detection limit. If your sample is from a soil or plant extraction, consider a more concentrated extraction to increase the amount of boron in the sample. If your samples have high ODs consider diluting them so they fall within the linear range of the standard curve.
This kit is optimized to detect boron in the form of borates and cannot be guaranteed to work with synthesized, and/or exotic boron compounds. However, samples may be treated to convert boron to borates.
For dry weight analysis, first completely dry the soil to remove all water weight. Next, we recommend mixing 5 g of soil (sieved to remove rocks and debris) with 50 mL of 50 mM HCl for 30 min. Centrifuge the sample to pellet precipitates and use the supernatant for the assay. Note: if there are soil particles floating make sure to avoid them when pipetting; your sample should be completely free of particulate matter.
For dry weight analysis, first completely dry the plant matter to remove all water weight. Next, we recommend burning 5 to 10 g of plant matter to ashes in a crucible at 500 degrees Fahrenheit for 3 hours in a drying oven. You may also use a blow torch to burn to ashes if a drying oven is not available. Mix all of the ashed plant material with 50 mL of 50 mM HCl for 30 min. Centrifuge the sample to pellet the ash and use the supernatant for the assay. Note: if there are ash particles floating make sure to avoid them when pipetting; your sample should be completely free of particulate matter.
If samples are strongly acidic or basic they should be adjusted to a pH of 5-7. Weak or dilute acids and bases do not need to be pH adjusted as the Working Reagent has sufficient buffering capacity.
In the [Boron] equation the ODBLANK is subtracted from the ODSAMPLE to remove any contribution from the reagent, plate, and reaction volume from the OD of the sample. ODH2O is subtracted from the ODSAMPLE BG so that only the background absorbance of the sample is further subtracted from the ODSAMPLE or, in other words, to make sure that the plate and reaction volume contributions to the ODSAMPLE are not subtracted out twice.
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