Difference between revisions of "Team:NCKU Tainan/Notebook Functional Test"

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#photo-left { background-image: url("/wiki/images/c/c1/T--NCKU_Tainan--sample2.jpg"); }
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            #photo-left { background-image: url("/wiki/images/c/cb/T--NCKU_Tainan--Functional.png"); }
</style>
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        </style>
<div class="container-fluid" style="margin-top:100px">
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        <div class="container-fluid" style="margin-top:100px">
<div class="head">NOTE / Functional Test</div>
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                <div class="head">NOTE / Functional Test</div>
<div class="content row">
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                <div class="content row">
<div class="col-md-9">
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                    <div class="col-md-9">
<div class="head2">Notebook - Functional Test</div>
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                        <div class="head2">Notebook - Functional Test</div>
<div class="title-line" id="sec1"></div>
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                        <div class="title-line long" id="sec1"></div>
<div class="title-content"></div>
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                        <div class="title-content">Compare promoter efficiency of Pcar with PI</div>
<h5></h5>
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                        <div class="text-content">
<p></p>
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                          <h5>May 14/2016</h5>
<h5></h5>
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                          <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-Pcar-RBS-amilCP-TT and pMD19T-PI-RBS-amilCP-TT from LB agar plate .</p>
<p></p>
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                          <h5>May 15/2016</h5>
<h5></h5>
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                          <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
<p></p>
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                          <h5>May 16/2016</h5>
<p></p>
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                          <p>Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM</p>
<h5></h5>
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                          <p>Readouts: measure OD 580 kinetically for 3.5 hr and compare the efficiency of Pcar and PI</p>
<p></p>
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                        </div>
<p></p>
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                        <div class="title-line long" id="sec2"></div>
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                        <div class="title-content">Glucose-Color detection test</div>
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                        <div class="text-content">
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                          <h5>May 20/2016</h5>
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                          <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate .</p>
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                          <h5>May 21/2016</h5>
 +
                          <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 +
                          <p>Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
 +
                          <p>Readouts: measure OD 580 kinetically for 3.5 hr.</p>
 +
                        </div>
  
</div>
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                        <div class="title-line long" id="sec3"></div>
<div class="col-md-3">
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                        <div class="title-content">Urine Glucose-Color detection test</div>
<ul id="sidemenu">
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                        <div class="text-content">
<li><a href="#" onclick="toEvent('section1');">Section 1</a></li>
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                          <h5>Jun. 17/2016</h5>
<li><a href="#" onclick="toEvent('section2');">Section 2</a></li>
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                          <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate .</p>
</ul>
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                          <h5>Jun. 18/2016</h5>
</div>
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                          <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
</div>
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                          <p>Sample preparation :collect urine sample from non-diabetic healthy individuals , and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.</p>
</div> <!-- /.container-fluid -->
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                          <p>Induction : add 100 μl of bacteria culture to 96 well plate and add Sample Urine to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
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                          <p>Readouts: measure OD 580 kinetically for 3.5 hr.</p>
 +
                        </div>
 +
 
 +
                        <div class="title-line long" id="sec4"></div>
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                        <div class="title-content">2-hour Glucose-Fluoresence detection test</div>
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                        <div class="text-content">
 +
                          <h5>Jul. 15/2016</h5>
 +
                          <p>Transformed plasmid (pMD19T-PpI-B0034-E1010) to BL21 competent cell.</p>
 +
                          <h5>Jul. 16/2016</h5>
 +
                          <p>Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one colony from the agar plate.</p>
 +
                          <h5>Jul. 17/2016</h5>
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                          <p>Extracted plasmid and confirmed plasmid size by gel electrophoresis.<br>
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                          <i class="glyphicon glyphicon-ok"></i>Result: The size of pMD19T-PI-RBS-mRFP was correct.</p>
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                          <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.</p>
 +
                          <h5>Jul. 18/2016</h5>
 +
                          <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP from LB agar plate .</p>
 +
                          <h5>Jul. 19/2016</h5>
 +
                          <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 +
                          <p>Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
 +
                          <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.</p>
 +
                        </div>
 +
 
 +
                        <div class="title-line long" id="sec5"></div>
 +
                        <div class="title-content">12-hour Glucose-Fluoresence detection test</div>
 +
                        <div class="text-content">
 +
                          <h5>Jul. 25/2016</h5>
 +
                          <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate .</p>
 +
                          <h5>Jul. 26/2016</h5>
 +
                          <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 +
                          <p>Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
 +
                          <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.</p>
 +
                        </div>
 +
 
 +
                        <div class="title-line long" id="sec6"></div>
 +
                        <div class="title-content">Lysis test</div>  
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                        <div class="text-content">
 +
                          <h5>Aug. 22/2016</h5>
 +
                          <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-P<sub>BAD</sub>-RBS-lysis-TT from LB agar plate .</p>
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                          <h5>Aug. 23/2016</h5>
 +
                          <p>Transfer : 20 ml of modified M9 (with Ampicilin) added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.</p>
 +
                          <p>Readouts: measure OD 600 kinetically with plate readerfor 4 hours .</p>
 +
                        </div>
 +
 
 +
                        <div class="title-line long" id="sec7"></div>
 +
                        <div class="title-content">Lysis test-2</div>
 +
                        <div class="text-content">
 +
                          <h5>Sep. 01/2016</h5>
 +
                          <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-P<sub>BAD</sub>-RBS-lysis-TT from LB agar plate .</p>
 +
                          <h5>Sep. 02/2016</h5>
 +
                          <p>Transfer : 20 ml of modified M9 (with Ampicilin)added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.</p>
 +
                          <p>Readouts: measure OD 600 kinetically with plate readerfor 4 hours .</p>
 +
                        </div>
 +
 
 +
                        <div class="title-line long" id="sec8"></div>
 +
                        <div class="title-content">2-hour Urine Glucose-Fluoresence detection test</div>
 +
                        <div class="text-content">
 +
                          <h5>Sep. 05/2016</h5>
 +
                          <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate .</p>
 +
                          <h5>Sep. 06/2016</h5>
 +
                          <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 +
                          <p>Sample preparation :collect urine sample from non-diabetic healthy individuals , and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.</p>
 +
                          <p>Induction : add 100 ul of bacteria culture to 96 well plate and add urine sample to final concentration : 0, 5, 15, 30, 60, 120 mM</p>
 +
                          <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.</p>
 +
                        </div>
 +
 
 +
                        <div class="title-line long" id="sec9"></div>
 +
                        <div class="title-content">12-hour Glucose-Fluoresence detection test-1 </div>
 +
                        <div class="text-content">
 +
                          <h5>Sep. 12/2016</h5>
 +
                          <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate .</p>
 +
                          <h5>Sep. 13/2016</h5>
 +
                          <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 +
                          <p>Sample preparation :collect urine sample from non-diabetic healthy individuals , and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.</p>
 +
                          <p>Induction : add 100 μl of bacteria culture to 96 well plate and add urine sample to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
 +
                          <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.</p>
 +
                        </div>
 +
                    </div>
 +
                </div>
 +
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Revision as of 01:37, 18 October 2016

Notebook Functional Test - iGEM NCKU

NOTE / Functional Test
Notebook - Functional Test
Compare promoter efficiency of Pcar with PI
May 14/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-Pcar-RBS-amilCP-TT and pMD19T-PI-RBS-amilCP-TT from LB agar plate .

May 15/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

May 16/2016

Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM

Readouts: measure OD 580 kinetically for 3.5 hr and compare the efficiency of Pcar and PI

Glucose-Color detection test
May 20/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate .

May 21/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM.

Readouts: measure OD 580 kinetically for 3.5 hr.

Urine Glucose-Color detection test
Jun. 17/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate .

Jun. 18/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Sample preparation :collect urine sample from non-diabetic healthy individuals , and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.

Induction : add 100 μl of bacteria culture to 96 well plate and add Sample Urine to final concentration : 0, 5, 15, 30, 60, 120 mM.

Readouts: measure OD 580 kinetically for 3.5 hr.

2-hour Glucose-Fluoresence detection test
Jul. 15/2016

Transformed plasmid (pMD19T-PpI-B0034-E1010) to BL21 competent cell.

Jul. 16/2016

Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one colony from the agar plate.

Jul. 17/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pMD19T-PI-RBS-mRFP was correct.

Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.

Jul. 18/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP from LB agar plate .

Jul. 19/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM.

Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.

12-hour Glucose-Fluoresence detection test
Jul. 25/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate .

Jul. 26/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM.

Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.

Lysis test
Aug. 22/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PBAD-RBS-lysis-TT from LB agar plate .

Aug. 23/2016

Transfer : 20 ml of modified M9 (with Ampicilin) added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.

Readouts: measure OD 600 kinetically with plate readerfor 4 hours .

Lysis test-2
Sep. 01/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PBAD-RBS-lysis-TT from LB agar plate .

Sep. 02/2016

Transfer : 20 ml of modified M9 (with Ampicilin)added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.

Readouts: measure OD 600 kinetically with plate readerfor 4 hours .

2-hour Urine Glucose-Fluoresence detection test
Sep. 05/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate .

Sep. 06/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Sample preparation :collect urine sample from non-diabetic healthy individuals , and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.

Induction : add 100 ul of bacteria culture to 96 well plate and add urine sample to final concentration : 0, 5, 15, 30, 60, 120 mM

Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.

12-hour Glucose-Fluoresence detection test-1
Sep. 12/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate .

Sep. 13/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Sample preparation :collect urine sample from non-diabetic healthy individuals , and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.

Induction : add 100 μl of bacteria culture to 96 well plate and add urine sample to final concentration : 0, 5, 15, 30, 60, 120 mM.

Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.