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| <div class="twelve columns centered text-center"> | | <div class="twelve columns centered text-center"> |
| <h1>Interlab</h1> | | <h1>Interlab</h1> |
− | <p class="title1" style="text-align:center">The Peking iGEM 2016 team is participating in the third year of the iGEM Interlab study along with about 100 teams.We focused on quantify expression of GFP in common, comparable or absolute units. </p> | + | <p class="title1" style="text-align:center">The Peking iGEM 2016 team is participating in the third year of the iGEM Interlab study along with about 100 teams. We focused on quantifying expression of GFP in common, comparable or absolute units. </p> |
| </div> | | </div> |
| </div> | | </div> |
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| o Test Device 2: J23106.B0034.E0040.B0015 in pSB1C3<br> | | o Test Device 2: J23106.B0034.E0040.B0015 in pSB1C3<br> |
| o Test Device 3: J23117.B0034.E0040.B0015 in pSB1C3<br> | | o Test Device 3: J23117.B0034.E0040.B0015 in pSB1C3<br> |
− | o Positive Control Device: I20270 in pSB1C3 Also located in Kit Plate 3, well 8P<br> | + | o Positive Control Device: I20270 in pSB1C3 (Also located in Kit Plate 3, well 8P)<br> |
− | o Negative Control Device: R0040 in pSB1C3 Also located in Kit Plate 2, well 6F<br> | + | o Negative Control Device: R0040 in pSB1C3 (Also located in Kit Plate 2, well 6F)<br> |
| </p> | | </p> |
| </div> | | </div> |
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| • LUDOX: one tube with 30% colloidal silica suspended in 1mL of water<br> | | • LUDOX: one tube with 30% colloidal silica suspended in 1mL of water<br> |
| • 1xPBS (phosphate buffered saline)<br> | | • 1xPBS (phosphate buffered saline)<br> |
− | • Terrific broth (at half strength: 0.5x TB) or can use LB (Luria Bertani) media as an alternative<br> | + | • LB (Luria Bertani) media<br> |
| • Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)<br> | | • Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)<br> |
| • 50 ml Falcon tube (or equivalent) or 250 ml shake flask for cell growth<br> | | • 50 ml Falcon tube (or equivalent) or 250 ml shake flask for cell growth<br> |
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| <div class="texttitle">Description</div> | | <div class="texttitle">Description</div> |
− | <img src="https://static.igem.org/mediawiki/2016/9/97/T--Peking--image_interlab_1.jpg"> | + | <img src="https://static.igem.org/mediawiki/2016/9/97/T--Peking--image_interlab_1.jpg"> |
− | <p style="text-align:center;"><b>Figure 1. From left to right:D1, D2, D3, Negative, Positive.</b></p>
| + | |
| <br> | | <br> |
| <img src="https://static.igem.org/mediawiki/2016/a/ab/T--Peking--image_interlab_2.jpg"> | | <img src="https://static.igem.org/mediawiki/2016/a/ab/T--Peking--image_interlab_2.jpg"> |
| + | <p style="text-align:center;"><b>Figure 1. Above: from left to right: D1, D2, D3, Negative, Positive. Below: five mediums containing five different bacteria.</b></p> |
| <br> | | <br> |
− | <p>Plasmids containing promoters and GFP were taken from The 2016 DNA Distribution Kit and all devices were transformed into E.coli. Fluorescence of colonies was checked up under UV light. </p> | + | <p>Plasmids containing promoters and GFP were taken from The 2016 DNA Distribution Kit and all devices were transformed into <i>E. coli</i>. Fluorescence of colonies was checked up under UV light. </p> |
| <p>5 ml of liquid LB-M medium with chloramphenicol were inoculated with two chosen colonies of each device. Liquid cultures were incubated for 16~18 hours in HONOUR INCURATOR SHAKER placed in incubator. OD of these cultures was measured by MAPADA UV-3100PC SPECTROPHOTOMETER and diluted to 0.02. Fluorescence of biological and also technical replicates was measured using Thermo VARIOSKAN FLASH following our protocol. | | <p>5 ml of liquid LB-M medium with chloramphenicol were inoculated with two chosen colonies of each device. Liquid cultures were incubated for 16~18 hours in HONOUR INCURATOR SHAKER placed in incubator. OD of these cultures was measured by MAPADA UV-3100PC SPECTROPHOTOMETER and diluted to 0.02. Fluorescence of biological and also technical replicates was measured using Thermo VARIOSKAN FLASH following our protocol. |
| </p> | | </p> |
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| <img src="https://static.igem.org/mediawiki/2016/0/04/T--Peking--image_interlab_7.png"> | | <img src="https://static.igem.org/mediawiki/2016/0/04/T--Peking--image_interlab_7.png"> |
| <p ><b>Table 5. Blank subtraction and correction of Abs600 measurement.</b></p> | | <p ><b>Table 5. Blank subtraction and correction of Abs600 measurement.</b></p> |
− | <img src="https://static.igem.org/mediawiki/2016/e/ee/T--Pek | + | |
| + | |
| + | |
| + | |
| + | |
| + | <img src="https://static.igem.org/mediawiki/2016/e/ee/T--Peking--image_interlab_8.png"> |
| + | <img src="https://static.igem.org/mediawiki/2016/7/71/T--Peking--image_interlab_9.png"> |
| + | <p ><b>Figure 3. Blank subtraction and correction of Abs600 measurement.</b></p> |
| + | <p ><b>Table 6. Raw data of fluorescence measurement.</b></p> |
| + | <img src="https://static.igem.org/mediawiki/2016/5/52/T--Peking--image_interlab_10.png"> |
| + | <p ><b>Table 7. Blank substraction and correction of fluorescence measurement.</b></p> |
| + | <img src="https://static.igem.org/mediawiki/2016/3/32/T--Peking--image_interlab_11.png"> |
| + | <img src="https://static.igem.org/mediawiki/2016/6/67/T--Peking--image_interlab_12.png"> |
| + | <p ><b>Figure 4. Blank subtraction and correction of fluorescence measurement.</b></p> |
| + | <p ><b>Table 8.Raw data of Fl/Abs600.</b></p> |
| + | <img src="https://static.igem.org/mediawiki/2016/d/d3/T--Peking--image_interlab_13.png"> |
| + | <p ><b>Table 9. Raw data of average and SD.</b></p> |
| + | <img src="https://static.igem.org/mediawiki/2016/a/a1/T--Peking--image_interlab_14.png"> |
| + | <img src="https://static.igem.org/mediawiki/2016/0/07/T--Peking--image_interlab_15.png"> |
| + | <p ><b>Figure 4. Average level of devices.</b></p> |
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| + | <div class="texttitle">Discussion</div> |
| + | <p>It is noticeable that the promoter of the Device 1 is strongest followed by the promoter of the Device 2 and Device 3.</p> |
| + | |
| + | <h3 class="classic-title";"><span>Appendix</span></h3> |
| + | <p>Individuals responsible for conducting InterLab study |
| + | • Dong Yiming measured the devices. |
| + | • Li Cheng processed the data. |
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| + | <script src='https://2016.igem.org/Template:Peking/Javascript/modernizr_js?action=raw&ctype=text/javascript'></script> |
| + | <script type='text/javascript' src='https://2016.igem.org/Template:Peking/Javascript/fjquery_polaroid?action=raw&ctype=text/javascript'></script> |
| + | <script type='text/javascript' src='https://2016.igem.org/Template:Peking/Javascript/fjquery_easing?action=raw&ctype=text/javascript'></script> |
| + | <script type='text/javascript' src='https://2016.igem.org/Template:Peking/Javascript/fjquery_transform_0_8_0_min?action=raw&ctype=text/javascript'></script> |
| + | <script type='text/javascript' src='https://2016.igem.org/Template:Peking/Javascript/fjquery_preloader?action=raw&ctype=text/javascript'></script> |
| + | <!--quotations from flexslider: end--> |
| + | |
| + | <!--quotations from black: start--> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_sticky?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_easing_1_3_pack?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/bootstrap_min?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_patallax_1_1_3?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/appear?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/modernizr?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_prettyPhoto?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/isotope?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_bxslider_min?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_cycle_all?action=raw&ctype=text/javascript" ></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/jquery_maximage?action=raw&ctype=text/javascript"></script> |
| + | <script type='text/javascript' src="https://2016.igem.org/Template:Peking/Javascript/scripts?action=raw&ctype=text/javascript "></script> |
| + | <!--quotations from black: end--> |
| + | |
| + | |
| + | </body> |