Difference between revisions of "Team:NTHU Formosa/InterLab"

 
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     <h2 class="w3-wide">IGEM 2018 Interlab Study</h2>
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<br><br><br>
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    <h3 class="w3-wide">Overall To-Do List </h3>
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     <h2 class="w3-wide" style="font-size:60px;font-family:Quicksand;">2018 iGEM Interlab Study</h2><br>
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    <p class="w3-justify">
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Calibration 1：​​ Reference point - LUDOX Protocol
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Record settings for standard measurements
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Record measurements
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Fill out corresponding Excel sheet
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Calibration 2：​Particle Standard Curve - Microsphere Protocol
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<p class="w3-center" style="font-size:40px;font-family:Quicksand;">Overall To-Do List</p><br>
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Prepare Microsphere Stock Solution
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<br>
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Serial dilution
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    <p class="w3-justify" style="font-size:25px;padding-left:150px;"><b>✔ Calibration 1：OD<sub>600</sub> Reference point - LUDOX Protocol</b></p>
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Measure Abs600 of all samples in instrument
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Record measurements
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    <p class="w3-justify" style="font-size:20px;padding-left:150px"><br></p>
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Fill out corresponding Excel sheet
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">1.Record settings for standard measurements</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">2.Record measurements</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">3.Fill out corresponding Excel sheet</p><br>
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    <p class="w3-justify" style="font-size:25px;padding-left:150px;"><b>✔ Calibration 2：​Particle Standard Curve - Microsphere Protocol</b></p>
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    <p class="w3-justify" style="font-size:20px;padding-left:150px"><br></p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">1.Prepare Microsphere Stock Solution</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">2.Serial dilution</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">3.Measure Abs600 of all samples in instrument</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">4.Record measurements</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">5.Fill out corresponding Excel sheet</p><br>
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    <p class="w3-justify" style="font-size:25px;padding-left:150px;"><b>✔ Calibration 3：Fluorescence standard curve - Fluorescein Protocol </b></p>
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    <p class="w3-justify" style="font-size:20px;padding-left:150px"><br></p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">1.Resuspend and dilute fluorescein</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">2.Serial dilutions</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">3.Measure fluorescence of all samples in instrument</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">4.Record measurements</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px;">5.Fill out corresponding Excel sheet</p><br>
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    <p class="w3-justify" style="font-size:25px;padding-left:150px"><b>✔ Cell Measurement protocol</b></p>
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<p class="w3-justify" style="font-size:20px;padding-left:150px"><br></p>
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    <p class="w3-justify" style="font-size:20px;padding-left:240px">Day 1: Transform E.Coli DH5α with plasmids (all in pSB1C3)</p>
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Calibration 3：Fluorescence standard curve - Fluorescein Protocol
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    <p class="w3-justify" style="font-size:20px;padding-left:240px">Day 2: Pick 2 colonies per plate (5 mL LB medium + Chloramphenicol.  </p>
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Resuspend and dilute fluorescein
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Serial dilutions
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    <p class="w3-justify" style="font-size:20px;padding-left:296px;">Grow 16-18 hours @ 37 C and 220 rpm</p>
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Measure fluorescence of all samples in instrument
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Record measurements
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Fill out corresponding Excel sheet
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Cell Measurement protocol
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    <p class="w3-justify" style="font-size:20px;padding-left:240px">Day 3:
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Day 1: Transform E.Coli DH5α with plasmids (all in pSB1C3)
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      Dilution cultures in LB+Chloramphenicol <br>
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Day 2: pick 2 colonies per plate (5 mL LB medium + Chloramphenicol. Grow 16-18 hours @ 37 C and 220 rpm
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      <p class="w3-justify" style="font-size:20px;padding-left:304px;">Measure OD<sub>600</sub><br></p>
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Day 3:
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      <p class="w3-justify" style="font-size:20px;padding-left:304px;">Further dilute<br></p>
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Dilution cultures in LB+Chloramphenicol  
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      <p class="w3-justify" style="font-size:20px;padding-left:304px;">Incubate<br></p>
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Measure  
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      <p class="w3-justify" style="font-size:20px;padding-left:304px;">Abs600 and fluorescence measurement<br></p>
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Further dilute
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      <p class="w3-justify" style="font-size:20px;padding-left:304px;">Record measurements<br></p>
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Incubate
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      <p class="w3-justify" style="font-size:20px;padding-left:304px;">Fill out corresponding Excel sheet<br></p>
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Abs600 and fluorescence measurement
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Record measurements
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Fill out corresponding Excel sheet
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    </p><br>
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    <p class="w3-justify" style="font-size:25px;padding-left:150px"><b>✔ Protocol：Colony Forming Units per 0.1 OD<sub>600</sub>  E.coli cultures</b></p>
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  <p class="w3-justify" style="font-size:20px;padding-left:150px"><br></p>
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      <p class="w3-justify" style="font-size:25px;padding-left:150px"><b>✔ To see the results, </b><a href="https://docs.google.com/spreadsheets/d/1GZ5xY4s5LGc7funlSDGDNl71Atd4TWjkDY37gka57aA/edit#gid=211336931"><src="https://docs.google.com/spreadsheets/d/1GZ5xY4s5LGc7funlSDGDNl71Atd4TWjkDY37gka57aA/edit#gid=211336931">click here</a>.</p>
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Protocol：Colony Forming Units per 0.1  E. coli cultures</p>
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Latest revision as of 14:10, 17 October 2018




2018 iGEM Interlab Study


Overall To-Do List



✔ Calibration 1：OD600 Reference point - LUDOX Protocol


1.Record settings for standard measurements

2.Record measurements

3.Fill out corresponding Excel sheet


✔ Calibration 2：​Particle Standard Curve - Microsphere Protocol


1.Prepare Microsphere Stock Solution

2.Serial dilution

3.Measure Abs600 of all samples in instrument

4.Record measurements

5.Fill out corresponding Excel sheet


✔ Calibration 3：Fluorescence standard curve - Fluorescein Protocol


1.Resuspend and dilute fluorescein

2.Serial dilutions

3.Measure fluorescence of all samples in instrument

4.Record measurements

5.Fill out corresponding Excel sheet


✔ Cell Measurement protocol


Day 1: Transform E.Coli DH5α with plasmids (all in pSB1C3)

Day 2: Pick 2 colonies per plate (5 mL LB medium + Chloramphenicol.

Grow 16-18 hours @ 37 C and 220 rpm

Day 3: Dilution cultures in LB+Chloramphenicol

Measure OD600

Further dilute

Incubate

Abs600 and fluorescence measurement

Record measurements

Fill out corresponding Excel sheet


✔ Protocol：Colony Forming Units per 0.1 OD600 E.coli cultures


✔ To see the results, click here.