Team:TU-Delft/FinalApplication
From 2013.igem.org
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<li>Dissolve the peptides. Signiferin was dissolved in water, in order to dissolve maximim 1M of NH4HCO3 was added in order to gain an end concentration of 5mM.</li> | <li>Dissolve the peptides. Signiferin was dissolved in water, in order to dissolve maximim 1M of NH4HCO3 was added in order to gain an end concentration of 5mM.</li> | ||
- | <li>The peptide solutions were measured, using the nanodrop, and we determined the initial absorbance. Using absorbance A= εcL=εc when L = 1cm [1] the concentration was measured</li> | + | <li>The peptide solutions were measured, using the nanodrop, and we determined the initial absorbance. Using absorbance A= εcL=εc when L = 1cm <a href="https://2013.igem.org/Team:TU-Delft/FinalApplication#references" style="text-decoration: none"">[1], [2]</a>the concentration was measured</li> |
<li> In order to make diffusion measurable, we folded the membrane like a filter paper and placed it in the peptide solution</li> | <li> In order to make diffusion measurable, we folded the membrane like a filter paper and placed it in the peptide solution</li> | ||
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on spectrophotometric determination of protein concentration</i>.[Online]. Available From: <a href=" http://classes.soe.ucsc.edu/bme220l/Spring11/Reading/Extinction-coefficients.pdf" style="text-decoration: none"" target="_blank">http://classes.soe.ucsc.edu/bme220l/Spring11/Reading/Extinction-coefficients.pdf</a> | on spectrophotometric determination of protein concentration</i>.[Online]. Available From: <a href=" http://classes.soe.ucsc.edu/bme220l/Spring11/Reading/Extinction-coefficients.pdf" style="text-decoration: none"" target="_blank">http://classes.soe.ucsc.edu/bme220l/Spring11/Reading/Extinction-coefficients.pdf</a> | ||
</li> | </li> | ||
- | <li> </li> | + | <li>C. N. Pace, F. Vajdos, L. Fee, G. Grimsley, and T. Gray,"How to measure and predict the molar absorption coefficient of a protein.", Protein Sci. 1995 November; 4(11): 2411–2423. </li> |
</ol> | </ol> | ||
Revision as of 15:13, 30 September 2013
BandAid Application
Band Aid is related to the final application of our project. Specifically, the idea is to use a band aid in which E.coli will be added. By locating the band aid on the wound, the S.aureus should be detected, the peptide will be produced and finally it will be pass through the band aid to the person who is infected.
Experiments
Aim: To check if it is feasible for the peptides to pass through the selected membranes (Cellulose Nitrate Membrane and PTFE Membrane).
Procedure:- Dissolve the peptides. Signiferin was dissolved in water, in order to dissolve maximim 1M of NH4HCO3 was added in order to gain an end concentration of 5mM.
- The peptide solutions were measured, using the nanodrop, and we determined the initial absorbance. Using absorbance A= εcL=εc when L = 1cm [1], [2]the concentration was measured
- In order to make diffusion measurable, we folded the membrane like a filter paper and placed it in the peptide solution
- Pierce Biotechnology: Extinction Coefficients, A guide to understanding extinction coefficients, with emphasis on spectrophotometric determination of protein concentration.[Online]. Available From: http://classes.soe.ucsc.edu/bme220l/Spring11/Reading/Extinction-coefficients.pdf
- C. N. Pace, F. Vajdos, L. Fee, G. Grimsley, and T. Gray,"How to measure and predict the molar absorption coefficient of a protein.", Protein Sci. 1995 November; 4(11): 2411–2423.
Absorbance(Maximim) | Absorbance(Signiferin) | Concentration in mM Maximim | Concentration in mM Signiferin | |
∅ | 0.51 | 0.74 | 0.34 | 0.49 |
t1 | 0 | 0 | 0 | 0 |
t2 | 0.03 | 0 | 0.02 | 0 |
t3 | 0.01 | 0.08 | 0.006 | 0.05 |
t20 | 0.02 | _ | 0.013 | - |
t60 | 0.16 | 0.27 | 0.1 | 0.18 |
t80 | 0.2 | 0.44 | 0.13 | 0.29 |
Table 2: PTFE Membrane, 0.1μm pore size
Absorbance(Maximim) | Absorbance(Signiferin) | Concentration in mM Maximim | Concentration in mM Signiferin | |
∅ | 0.51 | 0.74 | 0.02 | 0.49 |
t1 | 0 | 0 | 0 | 0 |
t2 | 0 | 0 | 0 | 0 |
t3 | 0 | 0 | 0 | 0 |
t20 | 0 | 0 | 0 | 0 |
t60 | 0 | 0 | 0 | 0 |
t80 | 0 | 0 | 0 | 0 |
Graphs
Conclusions