Difference between revisions of "Team:NYMU-Taipei/Composite Part"

 
(10 intermediate revisions by the same user not shown)
Line 3: Line 3:
 
<div id="page-content">
 
<div id="page-content">
  
 +
<img src="https://static.igem.org/mediawiki/2018/f/f3/T--NYMU-Taipei--composite-parts-cover.png" class="coverimage">
 +
<br>
 +
<br>
 +
<br>
  
<div class="column full_size judges-will-not-evaluate">
+
<div class="column">
<h3>★  ALERT! </h3>
+
<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>
+
<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>
+
</div>
+
  
 +
<div class="story">
 +
<div class="paragraphs">
  
<div class="clear"></div>
+
<h2>Our Favorite Composite Part: <a href="http://parts.igem.org/Part:BBa_K2751013">BBa_K2751013</a></h2>
 +
<h2>pCMV-ALB-mEGFP internal control</h2>
  
 +
<h3>Usage and Biology</h3>
  
 +
<p>This part contains the pCMV-ALB-mEGFP sequence, and is used as our internal control plasmid. The CMV promoter allows mEGFP to be expressed constantly when transfected into cells. Moreover, the signal peptide (ALB) allows mEGFP to be secreted extracellularly, so that signal detection can become much easier. </p>
  
 +
<img src="https://static.igem.org/mediawiki/parts/b/b8/T--NYMU-Taipei--IC1.png" style="width:900px;">
 +
<img src="https://static.igem.org/mediawiki/parts/0/09/T--NYMU-Taipei--IC2.png" style="width:500px;">
  
  
<div class="column full_size">
+
<h3>Characterization</h3>
<h1>Composite Parts</h1>
+
<p>We ran electrophoresis to check if the length of the part is correct. The length of CMV promoter, ALB sequence, and mEGFP sequence are 290bp, 56bp, and 720bp respectively. Therefore, we expect a product of 1066bp (1099bp along with a biobrick prefix). Gel photo is shown in the figure below.</p>
  
 +
<img src="https://static.igem.org/mediawiki/parts/4/46/T--NYMU-Taipei--parts-IC3.png" style="width:250px;">
  
<p>
+
<h3>Functionality Test</h3>
A composite part is a functional unit of DNA consisting of two or more basic parts assembled together. <a href="http://parts.igem.org/wiki/index.php/Part:BBa_I13507">BBa_I13507</a> is an example of a composite part, consisting of an RBS, a protein coding region for a red fluorescent protein, and a terminator.
+
<p>We transfected HEK293T cells via the GenJet Plus DNA transfection reagent and then collected cell supernatant for fluorescence measurement after 24 and 48 hours. Transparent alpha-MEM medium was used to minimize background noise. We used the Varioskan LUX spectral scanning multimode reader to measure fluorescence levels. Excitation wavelength was 488nm, emission wavelength was 509nm, and bandwidth was 5nm. Result is shown in the figure below. Blank medium control is taken from fresh medium, and cultured cell control is taken from the supernatant of cells that have not been transfected with the internal control plasmid. </p>
</p>
+
  
<p>New composite BioBrick devices can be made by combining existing BioBrick Parts (like Inverters, Amplifiers, Smell Generators, Protein Balloon Generators, Senders, Receivers, Actuators, and so on).</p>
 
</div>
 
  
<div class="column full_size">
+
<img src="https://static.igem.org/mediawiki/parts/8/8d/T--NYMU-Taipei--parts-IC4.png" style="width:500px;">
<div class="highlight decoration_background">
+
 
<h3>Note</h3>
+
<p>This page should list all the composite parts your team has made during your project. You must add all characterization information for your parts on the Registry. You should not put characterization information on this page. Remember judges will only look at the first part in the list for the Best Composite Part award, so put your best part first!</p>
+
 
</div>
 
</div>
 
</div>
 
</div>
  
 +
<div class="story">
 +
<div class="paragraphs">
  
 +
<h2>List of Composite Parts</h2>
  
<div class="column full_size">
+
<table style="width:100%">
<h3>Best Composite Part Special Prize</h3>
+
 
+
<p>To be eligible for this award, this part must adhere to <a href="http://parts.igem.org/DNA_Submission">Registry sample submission guidelines</a> and have been sent to the Registry of Standard Biological Parts. If you have a part you wish to nominate your team for this <a href="https://2018.igem.org/Judging/Awards">special prize</a>, make sure you add your part number to your <a href="https://2018.igem.org/Judging/Judging_Form">judging form</a> and delete the box at the top of this page.
+
 
+
<br><br>
+
<b>Please note:</b> Judges will only look at the first part number you list, so please only enter ONE (1) part number in the judging form for this prize. </p>
+
 
+
</div>
+
  
</html>
+
  <tr>
 +
    <th>Part Number</th>
 +
    <th>Description</th>
 +
    <th>Type</th>
 +
    <th>Length(bp)</th>
 +
  </tr>
 +
<tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2751013">BBa_K2751013</a></td>
 +
    <td>pCMV-ALB-mEGFP internal control</td>
 +
    <td>Composite</td>
 +
    <td>1066</td>
 +
  </tr>
 +
  <tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K27510">BBa_K2751009</a></td>
 +
    <td>LRP6BP1-YPet</td>
 +
    <td>Composite</td>
 +
    <td>1617</td>
 +
  </tr>
 +
<tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2751010">BBa_K2751010</a></td>
 +
    <td>Ubc9-YPet</td>
 +
    <td>Composite</td>
 +
    <td> 1305</td>
 +
  </tr>
 +
<tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2751012">BBa_K2751012</a></td>
 +
    <td>ALB signal peptide fused mCherry</td>
 +
    <td>Composite</td>
 +
    <td>793</td>
 +
  </tr>
 +
<tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2751014">BBa_K2751014</a></td>
 +
    <td>ALB-mEGFP</td>
 +
    <td>Composite</td>
 +
    <td>802</td>
 +
  </tr>
 +
 
 +
</table>

Latest revision as of 16:06, 16 October 2018




Our Favorite Composite Part: BBa_K2751013

pCMV-ALB-mEGFP internal control

Usage and Biology

This part contains the pCMV-ALB-mEGFP sequence, and is used as our internal control plasmid. The CMV promoter allows mEGFP to be expressed constantly when transfected into cells. Moreover, the signal peptide (ALB) allows mEGFP to be secreted extracellularly, so that signal detection can become much easier.

Characterization

We ran electrophoresis to check if the length of the part is correct. The length of CMV promoter, ALB sequence, and mEGFP sequence are 290bp, 56bp, and 720bp respectively. Therefore, we expect a product of 1066bp (1099bp along with a biobrick prefix). Gel photo is shown in the figure below.

Functionality Test

We transfected HEK293T cells via the GenJet Plus DNA transfection reagent and then collected cell supernatant for fluorescence measurement after 24 and 48 hours. Transparent alpha-MEM medium was used to minimize background noise. We used the Varioskan LUX spectral scanning multimode reader to measure fluorescence levels. Excitation wavelength was 488nm, emission wavelength was 509nm, and bandwidth was 5nm. Result is shown in the figure below. Blank medium control is taken from fresh medium, and cultured cell control is taken from the supernatant of cells that have not been transfected with the internal control plasmid.

List of Composite Parts

Part Number Description Type Length(bp)
BBa_K2751013 pCMV-ALB-mEGFP internal control Composite 1066
BBa_K2751009 LRP6BP1-YPet Composite 1617
BBa_K2751010 Ubc9-YPet Composite 1305
BBa_K2751012 ALB signal peptide fused mCherry Composite 793
BBa_K2751014 ALB-mEGFP Composite 802