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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    <h3 class="w3-wide">Overall To-Do List </h3>
 
  
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     <p class="w3-justify">
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     <p class="w3-center" style="font-size:60px;">IGEM 2018 Interlab Study</p>
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Calibration 1：​​ Reference point - LUDOX Protocol
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<br>
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Record settings for standard measurements
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<p class="w3-center" style="font-size:40px;">Overall To-Do List</p><br>
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Record measurements
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<br>
−
Fill out corresponding Excel sheet
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    <p class="w3-justify" style="font-size:30px;">Calibration 1：OD<sub>600</sub> Reference point - LUDOX Protocol</p>
 +
    <p class="w3-justify" style="font-size:20px;">1.Record settings for standard measurements</p>
 +
    <p class="w3-justify" style="font-size:20px;">2.Record measurements</p>
 +
    <p class="w3-justify" style="font-size:20px;">3.Fill out corresponding Excel sheet</p>
 +
    <p class="w3-justify" style="font-size:30px;">Calibration 2：​Particle Standard Curve - Microsphere Protocol</p>
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    <p class="w3-justify" style="font-size:20px;">1.Prepare Microsphere Stock Solution</p>
 +
    <p class="w3-justify" style="font-size:20px;">2.Serial dilution</p>
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    <p class="w3-justify" style="font-size:20px;">3.Measure Abs600 of all samples in instrument</p>
 +
    <p class="w3-justify" style="font-size:20px;">4.Record measurements</p>
 +
    <p class="w3-justify" style="font-size:20px;">5.Fill out corresponding Excel sheet</p>
 +
    <p class="w3-justify" style="font-size:30px;">Calibration 3：Fluorescence standard curve - Fluorescein Protocol </p>
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    <p class="w3-justify" style="font-size:20px;">1.Resuspend and dilute fluorescein</p>
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    <p class="w3-justify" style="font-size:20px;">2.Serial dilutions</p>
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    <p class="w3-justify" style="font-size:20px;">3.Measure fluorescence of all samples in instrument</p>
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    <p class="w3-justify" style="font-size:20px;">4.Record measurements</p>
 +
    <p class="w3-justify" style="font-size:20px;">5.Fill out corresponding Excel sheet</p>
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    <p class="w3-justify" style="font-size:40px;">Cell Measurement protocol</p>
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    <p class="w3-justify" style="font-size:25px;">Day 1:</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:70px">Transform E.Coli DH5α with plasmids (all in pSB1C3)</p>
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    <p class="w3-justify" style="font-size:25px;">Day 2:</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:70px">pick 2 colonies per plate (5 mL LB medium + Chloramphenicol. Grow 16-18 hours @ 37 C and 220 rpm</p>
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    <p class="w3-justify" style="font-size:25px;">Day 3:</p>
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    <p class="w3-justify" style="font-size:20px;padding-left:70px">
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      Dilution cultures in LB+Chloramphenicol <br>
 +
      Measure OD<sub>600</sub><br>
 +
      Further dilute<br>
 +
      Incubate<br>
 +
      Abs600 and fluorescence measurement<br>
 +
      Record measurements<br>
 +
      Fill out corresponding Excel sheet<br>
 +
     
 +
     
 +
     
 +
    </p>
 +
   
 +
   
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    <p class="w3-center" style="font-size:30px;">Protocol：Colony Forming Units per 0.1 OD<sub>600</sub>  E.coli cultures</p>
 +
 
  
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Calibration 2：​Particle Standard Curve - Microsphere Protocol
+
   
−
Prepare Microsphere Stock Solution
+
   
−
Serial dilution
+
 
−
Measure Abs600 of all samples in instrument
+
−
Record measurements
+
−
Fill out corresponding Excel sheet
+
−
 
+
−
Calibration 3：Fluorescence standard curve - Fluorescein Protocol
+
−
Resuspend and dilute fluorescein
+
−
Serial dilutions
+
−
Measure fluorescence of all samples in instrument
+
−
Record measurements
+
−
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
+
−
Further dilute
+
−
Incubate
+
−
Abs600 and fluorescence measurement
+
−
Record measurements
+
−
Fill out corresponding Excel sheet
+
−
 
+
−
Protocol：Colony Forming Units per 0.1  E. coli cultures</p>
+
  
 
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   </div>

Revision as of 14:41, 2 October 2018

IGEM 2018 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