Biomod/2011/TUM/TNT/LabbookA/2011/10/19: Difference between revisions
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=Camera calibration= | ==Camera calibration== | ||
In order to make | In order to make quantitative measurements of the distance of two flourophores we had to calibrate our fluorescence microscope. Therefor we took a picture of 5 scale bars (10 µm middle to middle). | ||
Insert: picture of scale bars | Insert: picture of scale bars | ||
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Calibration file: [[Image:Calibration_cam_green.xls]] | Calibration file: [[Image:Calibration_cam_green.xls]] | ||
==determined concentrations of labeled oligonucleotides== | |||
* staple 110 (helix 4) with ddUTP Atto 550: | |||
* staple 164 (helix 29) with ddUTP Atto 647N: 3.2µM | |||
==folded structure BM25_4/29== | |||
BM25_4/29 is intrinsically twisted, as BM24, but with fluorophors at staple 110 (helix 4, ddUTP Atto 550) and staple 164 (helix 29, dd UTP Atto 647N). <br> | |||
10µl each of both labeled oligonucleotides were added to the folding batch, replacing the usual 20µl ddH<sub>2</sub>O | |||
Revision as of 05:11, 20 October 2011
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Camera calibrationIn order to make quantitative measurements of the distance of two flourophores we had to calibrate our fluorescence microscope. Therefor we took a picture of 5 scale bars (10 µm middle to middle). Insert: picture of scale bars Hence the orientation isn't exactly vertically, we measured the horizontal shift of the bars to get the angle. With this we can calculate the right distance in pixels between the bars.
So our calibration is: Calibration file: File:Calibration cam green.xls determined concentrations of labeled oligonucleotides
folded structure BM25_4/29BM25_4/29 is intrinsically twisted, as BM24, but with fluorophors at staple 110 (helix 4, ddUTP Atto 550) and staple 164 (helix 29, dd UTP Atto 647N).
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