Difference between revisions of "Team:CSU CHINA/InterLab"

 
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<h1>Interlab</h1><br/>
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<div class="block main-text main-text" >
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<h class="highlight-title dark-blue"></br></br>Introduction</h>
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<p>
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Inerlab has come to the fifth session as a yardstick for measuring whether the standards of various laboratories around the world meet the requirements. Through standard curve measurement, 96-well plate readings and colony calculations, teams are able to ensure the measurement capabilities of each laboratory.  Interlab not only provides a basis for comparison between different laboratories, but also provides a basis for accurate research in synthetic biology.
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<h class="highlight-title dark-blue"></br>Materials</br></h>
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<h class="bold-text"></br></br>Plasmids</h>
  
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<p>
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Positive Control (BBa_I20270): well 2B <br/>
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Negative Control (BBa_R0040): well 2D <br/>
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<div class="column full_size judges-will-not-evaluate">
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Test Device 1 (BBa_J364000): well 2F <br/>
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<h3>★  ALERT! </h3>
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Test Device 2 (BBa_J364001): well 2H <br/>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</a>. </p>
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Test Device 3 (BBa_J364002): well 2J <br/>
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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://2018.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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Test Device 4 (BBa_J364007): well 2L <br/>
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Test Device 5 (BBa_J364008): well 2N <br/>
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Test Device 6 (BBa_J364009): well 2P
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</p><br/>
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<h class="bold-text"></br></br>Strains</h>
  
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<p>Escherichia coli strain DH5α</p><br/>
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<h class="bold-text"></br></br>Reagent</h>
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<p>
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1ml LUDOX<br/>
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ddH2O<br/>
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10ml 1X PBS<br/>
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LB (Luria Bertani) media<br/>

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Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)

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</p><br/>
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<h class="bold-text"></br></br>Instruments</h>
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<p>
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Pipettes<br/>
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96-well plate<br/>
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50 ml Falcon tube (or equivalent, preferably amber or covered in foil to block light)<br/>
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1.5-ml Eppendorf tubes for sample storage, ice bucket with ice<br/>
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Micropipettes and tips
<br/>
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</p>
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<h class="highlight-title dark-blue"></br></br>Methods</h>
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<p>
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There are 3 different parts of Interlab, including calibration, cell measurement and colony forming units. Calibrations are made up of OD6 00 Reference points, Particle Standard Curve, and Fluorescence standard curve, using LUDOX CL-X, Silica beads – Microsphere, Fluorescein provided in kit to generate a standardized curve. Cell measurement has 8 plasmids (Positive Control, Negative Control, Test Device 1, Test Device 2, Test Device 3, Test Device 4, Test Device 5, and Test Device 6) from Kit Plate 7 were transformed into E. coli DH5-alpha cells. After transformation and cultivation, the cells were used for experiments through the protocol. The third part is a procedure that aims to calibrate OD600 to colony forming unit (CFU) counts, which are directly relatable to the cell concentration of the culture.
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</p>
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<h class="highlight-title dark-blue"></br></br>Results</h>
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<img src="https://static.igem.org/mediawiki/2018/5/5f/T--CSU_CHINA--1_ODreferencePoint.png"/>
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<img src="https://static.igem.org/mediawiki/2018/thumb/3/3c/T--CSU_CHINA--interlab_OD600_2.jpg/800px-T--CSU_CHINA--interlab_OD600_2.jpg" width="400" height="300">
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<img src="https://static.igem.org/mediawiki/2018/a/a7/T--CSU_CHINA--2_ParticleStadardCurve.png"/>
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<img src="https://static.igem.org/mediawiki/2018/b/b1/T--CSU_CHINA--2_1GraphParticleStadardCurve.png"/>
  
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<img src="https://static.igem.org/mediawiki/2018/a/af/T--CSU_CHINA--2_2GraphParticleStadardCurve.png"/>
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<img src="https://static.igem.org/mediawiki/2018/6/68/T--CSU_CHINA--3_FluoresceinStandardCurve.png"/>
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<img src="https://static.igem.org/mediawiki/2018/4/44/T--CSU_CHINA--3_1GraphFluoresceinStandardCurve.png"/>
  
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<img src="https://static.igem.org/mediawiki/2018/c/c4/T--CSU_CHINA--3_2GraphFluoresceinStandardCurve.png"/>
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<img src="https://static.igem.org/mediawiki/2018/b/b7/T--CSU_CHINA--4_1RawPlateReaderMeasurementsFluorescence.png"/>
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<img src="https://static.igem.org/mediawiki/2018/d/d5/T--CSU_CHINA--4_2RawPlateReaderMeasurementsAbs600.png"/>
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<img src="https://static.igem.org/mediawiki/2018/0/00/T--CSU_CHINA--5_1FlorescencePerODfluorescein.png"/>
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<img src="https://static.igem.org/mediawiki/2018/2/2a/T--CSU_CHINA--5_1_1GraphuMFluoresceinOD600atSixHour.png"/>
  
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<img src="https://static.igem.org/mediawiki/2018/f/fa/T--CSU_CHINA--5_1_2GraphuMFluoresceinOD600atZeroHour.png"/>
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<h1>InterLab</h1>
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<br/>
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<h3>Bronze Medal Criterion #4</h3>
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<img src="https://static.igem.org/mediawiki/2018/f/f4/T--CSU_CHINA--5_2FlorescencePerODnetFluorescein.png"/>
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<p><b>Standard Tracks:</b> Participate in the Interlab Measurement Study and/or obtain new, high quality experimental characterization data for 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 2018 part number range.
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<br/>
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<br><br>
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<img src="https://static.igem.org/mediawiki/2018/d/de/T--CSU_CHINA--5_2_1GraphNetFluoresceinAUatZeroHour.png"/>
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For teams participating in the <a href="https://2018.igem.org/Measurement/InterLab">InterLab study</a>, all work must be shown on this page.  
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<img src="https://static.igem.org/mediawiki/2018/4/44/T--CSU_CHINA--5_2_2GraphNetFluoresceinAUatSixHour.png"/>
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<img src="https://static.igem.org/mediawiki/2018/6/63/T--CSU_CHINA--5_3FlorescencePerODAbs600.png"/>
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<img src="https://static.igem.org/mediawiki/2018/b/b6/T--CSU_CHINA--5_3_1GraphNetAbs600atZeroHour.png"/>
  
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<img src="https://static.igem.org/mediawiki/2018/5/51/T--CSU_CHINA--5_3_2GraphNetAbs600atSixHour.png"/>
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<img src="https://static.igem.org/mediawiki/2018/c/cc/T--CSU_CHINA--6_1FluorescencePerParticleMEFL.png"/>
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<img src="https://static.igem.org/mediawiki/2018/1/10/T--CSU_CHINA--6_1_1GraphMEFLParticleatSixHour.png"/>
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<img src="https://static.igem.org/mediawiki/2018/7/74/T--CSU_CHINA--6_1_2GraphMEFLParticleatZeroHour.png"/>
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<img src="https://static.igem.org/mediawiki/2018/9/99/T--CSU_CHINA--6_2FluorescencePerParticleNetFluorescein.png"/>
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<img src="https://static.igem.org/mediawiki/2018/c/c1/T--CSU_CHINA--6_3FluorescencePerParticleNetAbs600.png"/>
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<img src="https://static.igem.org/mediawiki/2018/5/55/T--CSU_CHINA--7_ColonyFormingUnits_.png"/>
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<br/>
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<img src="https://static.igem.org/mediawiki/2018/e/e9/T--CSU_CHINA--7_GraphColonyFormingUnits_.png"/>
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</center>
 
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{{CSU_CHINA/Footer}}

Latest revision as of 02:58, 18 October 2018


Interlab




Introduction

Inerlab has come to the fifth session as a yardstick for measuring whether the standards of various laboratories around the world meet the requirements. Through standard curve measurement, 96-well plate readings and colony calculations, teams are able to ensure the measurement capabilities of each laboratory. Interlab not only provides a basis for comparison between different laboratories, but also provides a basis for accurate research in synthetic biology.



Materials

Plasmids

Positive Control (BBa_I20270): well 2B
Negative Control (BBa_R0040): well 2D
Test Device 1 (BBa_J364000): well 2F
Test Device 2 (BBa_J364001): well 2H
Test Device 3 (BBa_J364002): well 2J
Test Device 4 (BBa_J364007): well 2L
Test Device 5 (BBa_J364008): well 2N
Test Device 6 (BBa_J364009): well 2P




Strains

Escherichia coli strain DH5α




Reagent

1ml LUDOX
ddH2O
10ml 1X PBS
LB (Luria Bertani) media

 Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)





Instruments

Pipettes
96-well plate
50 ml Falcon tube (or equivalent, preferably amber or covered in foil to block light)
1.5-ml Eppendorf tubes for sample storage, ice bucket with ice
Micropipettes and tips




Methods

There are 3 different parts of Interlab, including calibration, cell measurement and colony forming units. Calibrations are made up of OD6 00 Reference points, Particle Standard Curve, and Fluorescence standard curve, using LUDOX CL-X, Silica beads – Microsphere, Fluorescein provided in kit to generate a standardized curve. Cell measurement has 8 plasmids (Positive Control, Negative Control, Test Device 1, Test Device 2, Test Device 3, Test Device 4, Test Device 5, and Test Device 6) from Kit Plate 7 were transformed into E. coli DH5-alpha cells. After transformation and cultivation, the cells were used for experiments through the protocol. The third part is a procedure that aims to calibrate OD600 to colony forming unit (CFU) counts, which are directly relatable to the cell concentration of the culture.



Results