Objective
Learn how to maintain an OpenWetWare Notebook.
Samples
Sample 1
Mb lyophilized from Tris and KCl in water. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg myoglobin from Tris KCl
- OPD
- 2.6mg OPD in 2.6mL water
- H2O2
- 0.114mL 30% H2O2 in 0.887mL water
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: Mb_OPD_H2O2_tris_water_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 2
lyophilized from Tris and KCl in water. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin - no myoglobin this run
- 15mg lyophilized Tris KCl
- OPD
- 2.5mg OPD in 2.5mL water
- H2O2
- 0.114mL 30% H2O2 in 0.887mL water
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: KCl_OPD_H2O2_tris_water_25_SCANNUMBER.txt
- 0.225mL KCl solution added after 3 scans
- Should see this in the spectra
Sample 3
Mb lyophilized from Tris and KCl in methanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg myoglobin from Tris KCl in 1mL methanol
- OPD
- 2.4mg OPD in 2.4mL methanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL methanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: Mb_OPD_H2O2_tris_meoh_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 4
lyophilized Tris and KCl in methanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg Tris KCl in 1mL methanol
- OPD
- 2.5mg OPD in 2.5mL methanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL methanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: KCl_OPD_H2O2_tris_meoh_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 5
Mb lyophilized from Tris and KCl in ethanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg myoglobin from Tris KCl in 1mL ethanol
- OPD
- 3.7mg OPD in 3.7mL ethanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL ethanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: Mb_OPD_H2O2_tris_etoh_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 6
lyophilized Tris and KCl in ethanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg Tris KCl in 1mL ethanol
- OPD
- 3.4mg OPD in 3.4mL ethanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL ethanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: KCl_OPD_H2O2_tris_etoh_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 7
Mb lyophilized from Tris and KCl in isopropanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg myoglobin from Tris KCl in 1mL isopropanol
- OPD
- 3.0mg OPD in 3.0mL isopropanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL isopropanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: Mb_OPD_H2O2_tris_isop_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 8
lyophilized Tris and KCl in isopropanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg Tris KCl in 1mL isopropanol
- OPD
- 2.5mg OPD in 2.5mL isopropanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL isopropanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: KCl_OPD_H2O2_tris_isop_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 9
Mb lyophilized from Tris and KCl in 1-propanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg myoglobin from Tris KCl in 1mL 1-propanol
- OPD
- 2.9mg OPD in 2.9mL 1-propanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL 1-propanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: Mb_OPD_H2O2_tris_1prop_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 10
lyophilized Tris and KCl in 1-propanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg Tris KCl in 1mL 1-propanol
- OPD
- 4.0mg OPD in 4.0mL 1-propanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL 1-propanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: KCl_OPD_H2O2_tris_1prop_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 11
Mb lyophilized from Tris and KCl in 1-butanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg myoglobin from Tris KCl in 1mL 1-butanol
- OPD
- 2.7mg OPD in 2.7mL 1-butanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL 1-butanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: Mb_OPD_H2O2_tris_1but_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 12
lyophilized Tris and KCl in 1-butanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg Tris KCl in 1mL 1-butanol
- OPD
- 5.0mg OPD in 5.0mL 1-butanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL 1-butpanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: KCl_OPD_H2O2_tris_1but_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 13
Mb lyophilized from Tris and KCl in isobutanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg myoglobin from Tris KCl in 1mL isobutanol
- OPD
- 3.0mg OPD in 3.0mL isobutanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL isobutanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: Mb_OPD_H2O2_tris_isob_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
Sample 14
lyophilized Tris and KCl in isobutanol. scans every 10 seconds for 10 minutes. 1 scan = 1ms Integration, averaging over 200 scans
- Stock solutions:
- Myoglobin
- 15mg Tris KCl in 1mL isobutanol
- OPD
- 3.0mg OPD in 3.0mL isobutanol
- H2O2
- 0.114mL 30% H2O2 in 0.887mL isobutanol
- Experimental Run
- Sample chamber set to 25C and to stir
- N2 run through the cuvette holder to reduce condensation on cuvette
- 2.025mL OPD stock solution
- 0.75mL H2O2 stock solution
- Start Data Collection
- 1ms Integration
- 200 scan average
- Perform a scan every 10 seconds
- Stop after 10minutes
- Files saved as: KCl_OPD_H2O2_tris_isob_25_SCANNUMBER.txt
- 0.225mL Mb solution added after 3 scans
- Should see this in the spectra
- By the end of the day, the 30% peroxide that I had taken out of the fridge was showing bubbles. Concentration of peroxide should still be greater than that of OPD.
- Both the Mb in isobutanol and the KCl in isobutanol gave reddish solutions of roughly the same intensity at the end of the reaction. I will have to revisit these. Perhaps I'll do that tomorrow.
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