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− | <div class="vlogo"><img src="https://2017.igem.org/wiki/skins/Igem/images/IGEM_white_letters.png" alt="" /></div> | + | <div class="vlogo"> |
| + | <img src="https://2017.igem.org/wiki/skins/Igem/images/IGEM_white_letters.png" alt="" /> |
| + | </div> |
| <ul id="options" class="clearfix"> | | <ul id="options" class="clearfix"> |
| <li> | | <li> |
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| <div class="boxed"> | | <div class="boxed"> |
| <h2 class="header">COMMON PAGE</h2> | | <h2 class="header">COMMON PAGE</h2> |
| + | |
| + | <p > |
| + | <span style="font-weight:bold;font-size:22.0000pt;">The InterLab Study</span> |
| + | <span style="font-weight:bold;font-size:22.0000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="color:#FF0000;font-weight:bold;font-size:22.0000pt;font-family:'宋体';"></span> |
| + | <span style="color:#FF0000;font-weight:bold;font-size:22.0000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span> |
| + | <img src="https://static.igem.org/mediawiki/2017/3/32/T-ASWS-CHINA-lab2.png" height="196" width="550" /> |
| + | </span> |
| + | <span style="font-weight:bold;font-size:16.0000pt;"> </span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:16.0000pt;"> </span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:16.0000pt;"> </span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:16.0000pt;"> </span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:16.0000pt;"> </span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:16.0000pt;">Introduction </span> |
| + | <span style="font-weight:bold;font-size:16.0000pt;"></span> |
| + | </p> |
| <p> | | <p> |
− | Our idea of project comes from Hybridoma technology. We found some disadvantage about the old technology. In order to improve this technology, we started this project. We also want to know how most of people thinks about Hybridoma technology and our new approach. So we make a questioner to collect others view about our project. | + | Reliable and repeatable measurement is a key component to all engineering disciplines. The same holds true for synthetic biology, |
| + | which has also been called engineering biology. However, the ability to repeat measurements in different labs has been difficult. |
| </p> | | </p> |
− | <div class="images">
| |
− | <img src="https://static.igem.org/mediawiki/2017/e/e8/T-ASTWS-China-00001.jpeg" alt="" />
| |
− | </div>
| |
| <p> | | <p> |
− | To know the opinion from normal people, we started a questionnaire on WeChat. | + | This year is fourth InterLab study that organized by the iGEM Organization Measurement |
| + | Committee. It aims to establish a GFP measurement protocol based on engineering principles |
| + | that anyone with a plate reader can use in their lab and to improve measurement |
| + | tools and methods in laboratory work. |
| </p> | | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:16.0000pt;">E</span> |
| + | <span style="font-weight:bold;font-size:16.0000pt;font-family:'宋体';">xperiment </span> |
| + | <span style="font-weight:bold;font-size:16.0000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:14.0000pt;">The whole work flow:</span> |
| + | <span style="font-weight:bold;font-size:14.0000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <img src="https://static.igem.org/mediawiki/2017/3/34/T-ASWS-CHINA-lab4.jpg" height="262" width="560" /> |
| + | <span style="font-weight:bold;font-size:16.0000pt;"> </span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:14.0000pt;"> |
| + | The first day: Parts collection and transformation |
| + | </span> |
| + | <span style="font-weight:bold;font-size:14.0000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">Agents and </span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;">instruments that we used:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 1. |
| + | <a name="OLE_LINK4"></a> |
| + | <a name="OLE_LINK5"></a> |
| + | InterLab Measurement Kit |
| + | </span> |
| + | <span style="font-size:10.5000pt;">supplied by iGEM</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 2. Escherichia coli K-12 DH5α competent cells provided by ourselves |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">3. Water bath</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">4. Shake cultivation</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">5.</span> |
| + | <span style="color:#000000;font-size:10.5000pt;font-family:'Work Sans';background:#FFFFFF;"> </span> |
| + | <span style="font-size:10.5000pt;">Chloramphenicol as antibiotic</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 6. LB medium (liquid and agar plate) |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <a name="OLE_LINK1"></a> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"> |
| + | <a name="OLE_LINK2"></a> |
| + | P |
| + | </span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">rocedure:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">1. W</span> |
| + | <span style="font-size:10.5000pt;font-family:'宋体';">e </span> |
| + | <span style="font-size:10.5000pt;"> |
| + | transformed 8 parts listed in table 1 from the plate of InterLab Measurement Kit according to the given transformation protocol using Escherichia coli K-12 DH5α competent cells as required. |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <table style="border-collapse:collapse;width:247.8500pt;padding:0.0000pt 5.4000pt 0.0000pt 5.4000pt;"> |
| + | <tbody> |
| + | <tr> |
| + | <td width="132" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;font-family:'宋体';">Part name</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="113" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;font-family:'宋体';">Part ID</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="85" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;font-family:'宋体';">Location</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="132" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">Positive Control</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="113" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">BBa_I20270</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="85" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">21B</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="132" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">Negative Control</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="113" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">BBa_R0040</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="85" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">21D</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="132" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">Test Device 1</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="113" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">BBa_J364000</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="85" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">21F</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="132" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">Test Device 2</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="113" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">BBa_J364001</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="85" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">21H</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="132" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">Test Device 3</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="113" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">BBa_J364002</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="85" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">21J</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="132" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">Test Device 4</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="113" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">BBa_J364003</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="85" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">21L</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="132" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">Test Device 5</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="113" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">BBa_J364004</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="85" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">21N</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="132" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">Test Device 6</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="113" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">BBa_J364005</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | <td width="85" valign="top"> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">21P</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | </tbody> |
| + | </table> |
| + | <p > |
| + | <a name="OLE_LINK6"></a> |
| + | <span style="font-weight:bold;font-size:10.5000pt;">R</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">esults</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"> of transformation </span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p style="text-align:left;"> |
| + | <img src="https://static.igem.org/mediawiki/2017/2/28/T-ASWS-CHINA-lab3.png" height="283" width="554" /> |
| + | <span style="font-size:12.0000pt;font-family:'宋体';"> </span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:14.0000pt;"> |
| + | The second day: Colonies picking and cell growth |
| + | </span> |
| + | <span style="font-weight:bold;font-size:14.0000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <a name="OLE_LINK7"></a> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';"> |
| + | <a name="OLE_LINK8"></a> |
| + | Agents and |
| + | </span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;">instruments that we used:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">1. Shake cultivation</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">2.</span> |
| + | <span style="color:#000000;font-size:10.5000pt;font-family:'Work Sans';background:#FFFFFF;"> </span> |
| + | <span style="font-size:10.5000pt;">Chloramphenicol as antibiotic</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 3. LB medium (liquid and agar plate) |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">4. 8 plates with colonies</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;">P</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">rocedure:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">1. We p</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | ick 2 colonies from each of plate and inoculate it on 5-10 mL LB medium + Chloramphenicol. Grow the cells overnight (16-18 hours) at 37°C and 220 rpm. |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:14.0000pt;"> |
| + | The third day: Calibration, cell growth, sampling, and assay |
| + | </span> |
| + | <span style="font-weight:bold;font-size:14.0000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:12.0000pt;">Part I:</span> |
| + | <span style="font-weight:bold;font-size:12.0000pt;"> M</span> |
| + | <span style="font-weight:bold;font-size:12.0000pt;font-family:'宋体';">easurement of </span> |
| + | <span style="font-weight:bold;font-size:12.0000pt;">reference point of OD 600</span> |
| + | <span style="font-weight:bold;font-size:12.0000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">Agents and </span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;">instruments that we used:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">1. InterLab Measurement Kit</span> |
| + | <span style="font-size:10.5000pt;">supplied by iGEM</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 2. Microplate Reader: PerkinElmer Enspire 2300 |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">3. H</span> |
| + | <span style="font-size:10.5000pt;vertical-align:sub;">2</span> |
| + | <span style="font-size:10.5000pt;">0 </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 4. 96 well plate, black with flat, transparent. |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;">P</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">rocedure:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 1. Add 100 μl LUDOX into wells A1, B1, C1, D1 (or 1 mL LUDOX into cuvette) |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">2.</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Add 100 μl of H2O into wells A2, B2, C2, D2 (or 1 mL H2O into cuvette) |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">3.</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Measure absorbance 600 nm of all samples in all standard measurement modes in |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">Instrument.</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">4.</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Record the data in the table below or in your notebook |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">5</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Import data into Excel (OD600 reference point tab) |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;">R</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">esults</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"> of part I</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <table style="border-collapse:collapse;width:196.5000pt;padding:0.7500pt 0.7500pt 0.7500pt 0.7500pt;"> |
| + | <tbody> |
| + | <tr> |
| + | <td width="109" valign="bottom"> |
| + | <p > |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"> </span> |
| + | </p> |
| + | </td> |
| + | <td width="92" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">LUDOX-HS40</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td width="61" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">H2O</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="109" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">Replicate 1</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;background:#00CCFF;" width="92" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.047</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;background:#00CCFF;" width="61" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.037</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="109" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">Replicate 2</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;background:#00CCFF;" width="92" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.045</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;background:#00CCFF;" width="61" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.037</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="109" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">Replicate 3</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;background:#00CCFF;" width="92" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.047</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;background:#00CCFF;" width="61" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.037</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="109" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">Replicate 4</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;background:#00CCFF;" width="92" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.046</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;background:#00CCFF;" width="61" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.037</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="109" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">Arith. Mean</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;" width="92" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.04625</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;" width="61" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.037</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="109" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">Corrected Abs600</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;" width="92" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.00925</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td width="61" valign="bottom"> |
| + | <p > |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"> </span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="109" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">Reference OD600</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;" width="92" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">0.0425</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td width="61" valign="bottom"> |
| + | <p > |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"> </span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | <tr> |
| + | <td width="109" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">OD600/Abs600</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td style="border:0.5000pt solid #000000;" width="92" valign="bottom"> |
| + | <p style="text-align:left;vertical-align:;"> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';">4.594594595</span> |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"></span> |
| + | </p> |
| + | </td> |
| + | <td width="61" valign="bottom"> |
| + | <p > |
| + | <span style="color:#000000;font-size:12.0000pt;font-family:'宋体';"> </span> |
| + | </p> |
| + | </td> |
| + | </tr> |
| + | </tbody> |
| + | </table> |
| + | |
| + | <p > |
| + | <span style="font-weight:bold;font-size:12.0000pt;">Part II:</span> |
| + | <span style="font-weight:bold;font-size:12.0000pt;"> |
| + | Making the standard curve of fluorescein fluorescence |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">Agents and </span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;">instruments that we used:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">1. fluorescein (provided in kit)</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">2. 10ml 1xPBS </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 3. 96 well plate, black with flat, transparent. |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <a name="OLE_LINK10"></a> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"> |
| + | <a name="OLE_LINK11"></a> |
| + | P |
| + | </span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">rocedure:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;font-family:'宋体';">1. </span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Prepare the fluorescein stock solution as the |
| + | <a name="OLE_LINK9"></a> |
| + | protocol provided by iGEM. |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 2. Prepare the serial dilutions of fluorescein as the protocol. |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | <span style="font-size:10.5000pt;">3. </span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Repeat dilution series for rows B, C, D |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">4. </span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Measure fluorescence of all samples in all standard measurement modes in instrument |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">5. </span> |
| + | <span style="font-size:10.5000pt;">Record the data in your notebook</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">6. </span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Import data into Excel (fluorescein standard curve tab) |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;">R</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">esults</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"> of part II</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span> |
| + | <span> |
| + | <img src="https://static.igem.org/mediawiki/2017/e/e9/T-ASWS-CHINA-lab5.jpg" height="300" width="466" /> |
| + | </span> |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span> |
| + | <span> |
| + | <img src="https://static.igem.org/mediawiki/2017/d/d6/T-ASWS-CHINA-lab6.jpg" height="299" width="466" /> |
| + | </span> |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:12.0000pt;">Part II</span> |
| + | <span style="font-weight:bold;font-size:12.0000pt;font-family:'宋体';">I: Cell measurement</span> |
| + | <span style="font-weight:bold;font-size:12.0000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">Agents and </span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;">instruments that we used:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 1. Microplate Reader: PerkinElmer Enspire 2300 |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 2. Devices cultured in the first two days |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | 3. 96 well plate, black with flat, transparent. |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">4.</span> |
| + | <span style="font-size:10.5000pt;">50 mL falcon tube</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;">P</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">rocedure:</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">1.</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Set the instrument to read OD600 (as OD calibration setting) |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">2.</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Measure OD600 of the overnight cultures |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">3.</span> |
| + | <span style="font-size:10.5000pt;"> Record data</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">4.</span> |
| + | <span style="font-size:10.5000pt;"> Import data into Excel</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">5.</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Dilute the cultures to a target OD600 of 0.02 in 12 ml LB medium + Chloramphenicol |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | in 50 mL falcon tube (amber, or covered with foil to block light). |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">6.</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Incubate the cultures at 37°C and 220 rpm. |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">8.</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Take 500 μL samples of the cultures at 0, 2, 4, and 6 hours of incubation. (At each time |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;"> |
| + | point, we took a sample from each of the 8 devices, two colonies |
| + | per device, for a total of 16 samples per time point) |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">9.</span> |
| + | <span style="font-size:10.5000pt;"> Place samples on ice.</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">10.</span> |
| + | <span style="font-size:10.5000pt;"> |
| + | Measure samples at the end of sampling point you need to (OD and Fl measurement). |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">11.</span> |
| + | <span style="font-size:10.5000pt;"> Record data in the notebook</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-size:10.5000pt;">12.</span> |
| + | <span style="font-size:10.5000pt;"> Import data into Excel.</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span style="font-weight:bold;font-size:10.5000pt;">R</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;font-family:'宋体';">esults</span> |
| + | <span style="font-weight:bold;font-size:10.5000pt;"> of part III</span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span> |
| + | <span> |
| + | <img src="https://static.igem.org/mediawiki/2017/a/ab/T-ASWS-CHINA-lab7.jpg" height="366" width="522" /> |
| + | </span> |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | <p > |
| + | <span> |
| + | <span> |
| + | <img src="https://static.igem.org/mediawiki/2017/8/8b/T-ASWS-CHINA-lab8.jpg" height="362" width="522" /> |
| + | </span> |
| + | </span> |
| + | <span style="font-size:10.5000pt;"></span> |
| + | </p> |
| + | |
| </div> | | </div> |
| </article> | | </article> |