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The absorbance value was measured at 750 nm using visible spectrophotometer against distilled water blank. .
Track 1: 35S-GUS positiveositive control); track 2 6: pSH4-GUS transgenic plants; track 7: Nipponbare (negative control); track 8: water (blank control).
The absorbance of each dilution was measured at 520 and 700 nm against a distilled water blank using a UV-visible spectrophotometer (Specord 205 by Analytik Jena) and 1 cm path length glass cells.
After that, approximately 0.5 mL of the sample solution was transferred into a tube while 0.5 mL of water (blank test) was transferred into a second tube and 0.5 mL of proline standard solution were taken into three other tubes.
Detection limits ranged from 0.066 to 0.563 μg/L in water (mean of 0.202±0.141μg/L); meanwhile, noise and signal height ranged from 86 to 520 (mean of 205±126) and from 1526 to 5116 (mean of 3411±997), respectively, for 2 μg/L of fortified matrix water blank.
The amount of CO2 emitted from the respiration of soil sample and adsorbed by NaOH solution was determined every 3 4 days (longer incubation periods could result in anaerobic conditions) by back titration with 0.05 M HCl after addition of 7 mL of 0.5 M BaCl2 in both water blank and NaOH solutions.
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Water blanks were subtracted, and analyses conducted in triplicate.
Further, routine fortified ultrapure water blanks were analyzed along with the samples to check for any contamination.
Negative controls (water blanks) were included on each plate.
Multiple negative water blanks were tested, and a calibration curve was determined in parallel with each analysis.
Water blanks were included with each assay.
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