in the experiment to quantify the ability to produce siderophore in rice roots by CAS reagent. We have performed 3 times at different levels, but the OD measurement result is still negative. Tell me the reason
First time: 600ul bacterial epidemicin, 600ul CAS reagent. Measured at 630nm
Second time: 600ul bacterial epidemicin, 600ul CAS reagent. Measured at 530nm
3rd time: 900ul of bacterial epidemicin, 100ul of CAS reagent. Measured at 630nmm
Dear Hoa Ngọc Nguyễn
If OD measurement results for quantifying siderophore production in rice roots are consistently negative, there are several potential reasons. These include insufficient siderophore production, an ineffective CAS reagent, incorrect measurement parameters, experimental conditions, and technical errors. Insufficient siderophore production can occur due to the bacteria's inability to produce siderophores, while ineffective CAS reagents may not be suitable for detecting specific siderophores. The optimal wavelength for measuring OD should be appropriate for the CAS reagent and validated for siderophore quantification. Experimental conditions, such as pH, temperature, incubation time, and bacterial growth stage, can also impact siderophore production.
Technical errors, such as inaccurate pipetting or contamination, can lead to inconsistent or inaccurate results. To troubleshoot the issue, verify the bacterial strain, check the compatibility and specificity of the CAS reagent with the siderophores produced by the strain, validate measurement parameters, evaluate and optimize experimental conditions, review the experimental procedure, and consult relevant literature or experts for guidance. By systematically assessing these factors, potential causes for negative results can be identified and adjustments can be made to improve the experiment.
Some references:
Layer plate CAS assay for the quantitation of siderophore production and determination of exudation patterns for fungi Megan Y Andrews et al. J Microbiol Methods. 2016 Feb.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3577196/ TITLE: Use of Blue Agar CAS Assay for Siderophore Detection - PMC CONTENT: Journal List J Microbiol Biol Educ v.12(1); 2011 PMC3577196 J Microbiol Biol Educ. 2011; 12(1): 51–53.
- Badges
- Science method