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| </p> | | </p> |
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− | <div class="accordion" style="margin-top:100px"> | + | <div class="accordion" style="margin-top: 10px;"> |
| <div class="accordion-section"> | | <div class="accordion-section"> |
| <a class="accordion-section-title" href="#accordion-1">Spot analysis by hand: A step by step guidance</a> | | <a class="accordion-section-title" href="#accordion-1">Spot analysis by hand: A step by step guidance</a> |
| <div id="accordion-1" class="accordion-section-content" style="display:none; padding:15px;"> | | <div id="accordion-1" class="accordion-section-content" style="display:none; padding:15px;"> |
| + | <p> This step by step guidance shows you how to dertermine endpoint intensities of the binding experiment as shown above. Have a look at our <a href="https://2015.igem.org/Team:Freiburg/Project/iRIf">iRIF</a> page to getall the background informations about the detection method.</p> |
| <ol> | | <ol> |
| <li> During your iRIF experiment you take two pictures. One before the binding event and one after the binding event. Open both pictures in <a href="http://imagej.nih.gov/ij/">ImageJ</a>. </li> | | <li> During your iRIF experiment you take two pictures. One before the binding event and one after the binding event. Open both pictures in <a href="http://imagej.nih.gov/ij/">ImageJ</a>. </li> |
| <li> Go to "Process" and click on "Image Calculator..." </li> | | <li> Go to "Process" and click on "Image Calculator..." </li> |
− | <li> Set "Image1" to the image after the binding event and "Image2" to the image before the binding event. Choose "divide" as operation. Activate the "32-bit (float) result" and click ok. A new quotient picture will be calculated and opened. </li> | + | <li> Set "Image1" to the image after the binding event and "Image2" to the image before the binding event. Choose "divide" as operation. Activate the "32-bit (float) result" and click "ok". A quotient picture will be calculated and opened. </li> |
| <li> Go to "Image" then "Adjust" and klick on "Brightness/Contrast..." and play around with "brightness" and "contrast" till the spots get visible.</li> | | <li> Go to "Image" then "Adjust" and klick on "Brightness/Contrast..." and play around with "brightness" and "contrast" till the spots get visible.</li> |
− | <li> Mark the spots using the "oval" selection tool and press "T" on your keybord. Do this for every spot. Do not foret to make a selection of the background. <li> | + | <li> Mark the spots using the "oval" selection tool and press "T" on your keybord. Do this for every spot. Do not foret to make a selection of the background. </li> |
| <li> Go to the ROI-manager that poped up when you first pushed T and click on "Deselect" and then on "Measure". The results of your Measurment will pop up. You can save them in Excel.</li> | | <li> Go to the ROI-manager that poped up when you first pushed T and click on "Deselect" and then on "Measure". The results of your Measurment will pop up. You can save them in Excel.</li> |
| </ol> | | </ol> |