User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/21: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Meeting, Absorbance Data</span> | ||
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Absorbance spectra, especially those from the silica-coated AuNPs made with MHA and 5uL TES, are consistent with those from the literature (according to [http://pubs.acs.org/doi/full/10.1021/la9601871 Synthesis of Nanosized Gold−Silica Core−Shell Particles]). The silica-coated AuNPs show a peak at approximately 525nm.<br.> | Absorbance spectra, especially those from the silica-coated AuNPs made with MHA and 5uL TES, are consistent with those from the literature (according to [http://pubs.acs.org/doi/full/10.1021/la9601871 Synthesis of Nanosized Gold−Silica Core−Shell Particles]). The silica-coated AuNPs show a peak at approximately 525nm.<br.> | ||
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Since the AuNPs@SiO<sub>2</sub> synthesized with MHA and 5uL TES produced the best spectra, I think I will move onto the DNA attachment procedure using those | Since the AuNPs@SiO<sub>2</sub> synthesized with MHA and 5uL TES produced the best spectra, I think I will move onto the DNA attachment procedure using those. Additionally, I'm interested in varying the volume of TES added to see if higher or lower volumes will produce thicker or thinner silica-coatings, respectively (keeping the MHA constant and trying 10uL, 15uL, 20uL). I might also look at the volumes of TES used without MHA: since the higher volume of TES (40uL) did not seem to produce the expected spectra and the lower volume (20uL) did, I'd be interested to see the results of using 30uL, 15uL, 10uL, and 5uL of TES would be.<br.> | ||
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A final important note- PBS was used for ALL dilutions in this procedure, and sample were sonicated after dilution before being measured. The salt content of the PBS prevents the nanoparticles from aggregating. | A final important note- PBS was used for ALL dilutions in this procedure, and sample were sonicated after dilution before being measured. The salt content of the PBS prevents the nanoparticles from aggregating. | ||
===DNA/ThT Solutions=== | |||
Solutions of 100nM, 200nM and 700nM DNA with 2.5uM ThT were prepared and their fluorescence measured to make a more complete calibration curve (see [http://openwetware.org/wiki/User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/06/20 06/20/2013]). Fluorescence for the 200nM sample observed as expected, but the 100nM was much lower than expected and the 700nM much higher than expected. This inconsistency may be due to incorrect calibration of pipetman. Samples will be made again and run next week. | |||
Latest revision as of 22:46, 26 September 2017
 Meeting, Absorbance Data
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June 21, 2013Notes from Meeting
Absorbance Data
NotesAbsorbance spectra, especially those from the silica-coated AuNPs made with MHA and 5uL TES, are consistent with those from the literature (according to Synthesis of Nanosized Gold−Silica Core−Shell Particles). The silica-coated AuNPs show a peak at approximately 525nm.<br.> <br.> Since the AuNPs@SiO2 synthesized with MHA and 5uL TES produced the best spectra, I think I will move onto the DNA attachment procedure using those. Additionally, I'm interested in varying the volume of TES added to see if higher or lower volumes will produce thicker or thinner silica-coatings, respectively (keeping the MHA constant and trying 10uL, 15uL, 20uL). I might also look at the volumes of TES used without MHA: since the higher volume of TES (40uL) did not seem to produce the expected spectra and the lower volume (20uL) did, I'd be interested to see the results of using 30uL, 15uL, 10uL, and 5uL of TES would be.<br.> <br.> A final important note- PBS was used for ALL dilutions in this procedure, and sample were sonicated after dilution before being measured. The salt content of the PBS prevents the nanoparticles from aggregating. DNA/ThT SolutionsSolutions of 100nM, 200nM and 700nM DNA with 2.5uM ThT were prepared and their fluorescence measured to make a more complete calibration curve (see 06/20/2013). Fluorescence for the 200nM sample observed as expected, but the 100nM was much lower than expected and the 700nM much higher than expected. This inconsistency may be due to incorrect calibration of pipetman. Samples will be made again and run next week.
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