<p>This page is used by the judges to evaluate your team for the <a href="https://2017.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2017.igem.org/Judging/Awards"> award listed above</a>. </p>
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<ul class="page__drawer-list">
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2017.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
<p><b>Standard Tracks:</b> Participate in the Interlab Measurement Study and/or improve the characterization of an existing BioBrick Part or Device and enter this information on that part's Main Page in the Registry. The part that you are characterizing must NOT be from a 2017 part number range.
For teams participating in the <a href="https://2017.igem.org/Competition/InterLab_Study">InterLab study</a>, all work must be shown on this page.
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</h2>
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<p class="mdc-typography--body2">
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Many teachers and iGEM Headquarters says that reliable and repeatable measurement is a key component to all engineering disciplines. But to be honest it's difficult for laboratories all around the world to measure something in the same standard, so iGEM developing a robust measurement procedure for green fluorescent protein (GFP) every years to lead the team all over world could measure it.
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</p>
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<p class="mdc-typography--body2">
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It's the fourth year for iGEM to require teams which participate in finishing the interlab work. And GFP is a most widely used.
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</p>
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<p class="mdc-typography--body2">
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"All of the 2017 iGEM teams are invited and encouraged to participate in the Fourth International InterLaboratory Measurement Study in synthetic biology. We're hoping this study will get you excited for iGEM and help prepare you for the summer!" says by iGEM Headquarters. Actually, for us the interlab work not only to make fun but the more important part is that it can lead different laboratory to normalize their measure methods and machines so that we can use data from other lab easily, which will benefit our experiment and project a lot.
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</p>
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<p class="mdc-typography--body2">
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We all glad to participate in such significant international work with those friendly workmates all around world.
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</p>
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</p>
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<h2 class="mdc-typography--headline">Materials and methods</h2>
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</div>
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<h3 class="mdc-typography--title">Materials</h3>
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<ul class="page__article__list">
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<li>Plasmid DNA (1ng in total, 100 pg/uL in 10uL of ddH20)
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<ul>
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<li>Positive control BBa_I20270</li>
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<li>Negative control BBa_R0040</li>
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<li>Test Device 1: J23101.BCD2.E0040.B0015</li>
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<li>Test Device 2: J23106.BCD2.E0040.B0015</li>
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<li>Test Device 3: J23117.BCD2.E0040.B0015</li>
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<li>Test Device 4: J23101+I13504</li>
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<li>Test Device 5: J23106+I13504</li>
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<li>Test Device 6: J23117+I13504</li>
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</ul>
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</li>
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<li>Strain Used
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<ul>
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<li>Escherichia coli DH5α</li>
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</ul>
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</li>
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<li>Media Used
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<ul>
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<li>LB (Luria Bertani) media</li>
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</ul>
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</li>
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<li>Reagent Used
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<ul>
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<li>1xPBS (phosphate buffered saline)</li>
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<li>Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)</li>
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<li>FITC Standard: one tube with dried down FITC for creating a FITC standard</li>
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<li>LUDOX: one tube with 30% colloidal silica suspended in 1mL of water</li>
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</ul>
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</li>
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<li>Consumable Items
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<ul>
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<li>50 ml Falcon tube</li>
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<li>1.5 ml eppendorf tubes for sample storage</li>
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<li>Ice box with ice</li>
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<li>Pipettes and tip</li>
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<li>96 well plate</li>
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</ul>
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</li>
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</ul>
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<h3 class="mdc-typography--title">Machines</h3>
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<ul class="page__article__list">
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<li>
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<b>Thermo scientific</b>
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Varioskan Flash
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</li>
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<li>
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<b>CRYSTAL</b>
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Incubator Shaker
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</li>
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<li>
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<b>Yiheng-China</b>
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HZQ-F160A constant temperature incubator
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</li>
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<li>
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<b>AIRTECH</b>
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Vertical Flow Clean Bench
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</li>
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</ul>
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<h3 class="mdc-typography--title">Methods</h3>
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<ul class="page__article__list">
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<li>Calibration
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<ul>
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<li>OD600 Reference poin</li>
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<li>FITC fluorescence standard curve</li>
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</ul>
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</li>
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<li>Cell measurement
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<ul>
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<li>Transformation</li>
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<li>Measurements</li>
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</ul>
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</li>
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</ul>
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<h1 class="mdc-typography--display1">Data and Analysis</h1>
At first time, when we decide to transform plasmid into E.coli, what interesting is that we find device 1 can't growth in 170ug/ml or in 90ug/ml chloramphenicol at the same time we know exactly this plasmid is a high express one. Based on figure 3, we find that the growth curve of E.coli (except device 1) are all similar to the front part of S-Curve of Logistic regression, but it was strange that the two colony of device 1 seems don't change obviously in bacterial density which is different with other colonies. We conjectured this phenomenon came from the extremely high expression quantity of GFP which already produce severe cytotoxicity. As a result the growth of device 1 is very slow and can't growth in high concentration of chloramphenicol.
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</p>
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<p class="mdc-typography--body2">
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In figure 4 and figure 5, we analyzed the relationship between fluorescence and Abs 600. Then find that the promotor of GFP in device 1 actually is strongest in 8 plasmid which as 5 times as positive control, device 2 and device 4. The expression of GFP in device 5 seems very low, only one-tenth of positive control.
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</p>
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<p class="mdc-typography--body2">
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Also, we find that if the sampling time overlong, the E.coli growth will be restrain because of long time of low-temperature stress, in this suitcase, the growth of bacteria density is very slow and unparalleled in two colonies.
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</p>
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</article>
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</main>
</html>
</html>
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{{LanzhouFooter}}
Revision as of 09:53, 8 October 2017
Lanzhou
Lanzhou2017
Introduction
Background
Many teachers and iGEM Headquarters says that reliable and repeatable measurement is a key component to all engineering disciplines. But to be honest it's difficult for laboratories all around the world to measure something in the same standard, so iGEM developing a robust measurement procedure for green fluorescent protein (GFP) every years to lead the team all over world could measure it.
It's the fourth year for iGEM to require teams which participate in finishing the interlab work. And GFP is a most widely used.
"All of the 2017 iGEM teams are invited and encouraged to participate in the Fourth International InterLaboratory Measurement Study in synthetic biology. We're hoping this study will get you excited for iGEM and help prepare you for the summer!" says by iGEM Headquarters. Actually, for us the interlab work not only to make fun but the more important part is that it can lead different laboratory to normalize their measure methods and machines so that we can use data from other lab easily, which will benefit our experiment and project a lot.
We all glad to participate in such significant international work with those friendly workmates all around world.
Materials and methods
Materials
Plasmid DNA (1ng in total, 100 pg/uL in 10uL of ddH20)
Positive control BBa_I20270
Negative control BBa_R0040
Test Device 1: J23101.BCD2.E0040.B0015
Test Device 2: J23106.BCD2.E0040.B0015
Test Device 3: J23117.BCD2.E0040.B0015
Test Device 4: J23101+I13504
Test Device 5: J23106+I13504
Test Device 6: J23117+I13504
Strain Used
Escherichia coli DH5α
Media Used
LB (Luria Bertani) media
Reagent Used
1xPBS (phosphate buffered saline)
Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)
FITC Standard: one tube with dried down FITC for creating a FITC standard
LUDOX: one tube with 30% colloidal silica suspended in 1mL of water
Consumable Items
50 ml Falcon tube
1.5 ml eppendorf tubes for sample storage
Ice box with ice
Pipettes and tip
96 well plate
Machines
Thermo scientific
Varioskan Flash
CRYSTAL
Incubator Shaker
Yiheng-China
HZQ-F160A constant temperature incubator
AIRTECH
Vertical Flow Clean Bench
Methods
Calibration
OD600 Reference poin
FITC fluorescence standard curve
Cell measurement
Transformation
Measurements
Data and Analysis
Normalization work
Cell Measurement
Discussion
At first time, when we decide to transform plasmid into E.coli, what interesting is that we find device 1 can't growth in 170ug/ml or in 90ug/ml chloramphenicol at the same time we know exactly this plasmid is a high express one. Based on figure 3, we find that the growth curve of E.coli (except device 1) are all similar to the front part of S-Curve of Logistic regression, but it was strange that the two colony of device 1 seems don't change obviously in bacterial density which is different with other colonies. We conjectured this phenomenon came from the extremely high expression quantity of GFP which already produce severe cytotoxicity. As a result the growth of device 1 is very slow and can't growth in high concentration of chloramphenicol.
In figure 4 and figure 5, we analyzed the relationship between fluorescence and Abs 600. Then find that the promotor of GFP in device 1 actually is strongest in 8 plasmid which as 5 times as positive control, device 2 and device 4. The expression of GFP in device 5 seems very low, only one-tenth of positive control.
Also, we find that if the sampling time overlong, the E.coli growth will be restrain because of long time of low-temperature stress, in this suitcase, the growth of bacteria density is very slow and unparalleled in two colonies.
