Team:TU Eindhoven/Project/protocols

From 2014.igem.org

(Difference between revisions)
(Created page with "{{:Team:TU_Eindhoven/Template:NavPanel}} <html> <head> <title>Project Description</title> <meta http-equiv="Content-Type" content="text/html; charset=utf-8" /> <meta name="vie...")
 
(35 intermediate revisions not shown)
Line 42: Line 42:
      <p class="para">For this year's iGEM competition numerous protocols were devoloped to guide our experiments and keep our documentation neat and tidy. Because these protocols can also be useful to other projects, we decided to publish them on our wiki. You can find information and download links on this page down below.
      <p class="para">For this year's iGEM competition numerous protocols were devoloped to guide our experiments and keep our documentation neat and tidy. Because these protocols can also be useful to other projects, we decided to publish them on our wiki. You can find information and download links on this page down below.
-
<br><br>
+
<br>
-
To create such resilient bacteria the 2014 team has designed a ‘plug and play’ system using copper free click chemistry to attach different chemical groups to create bio-layers on E. Coli cell membranes. Circularly permuted OmpX (CPX), an outer membrane protein, was mutated to contain an azido-functionalized unnatural amino acid. CPX  functions as an anchor for any DBCO functionalized molecule to click onto. The polymers used in this project were designed to form hydrogels, which enables the bacteria to have antifouling properties.
+
 
   </p>
   </p>
Line 57: Line 57:
<div class="cont-grid_left">
<div class="cont-grid_left">
      <div class="Jasper">
      <div class="Jasper">
-
  <span>Application</span>  
+
  <span>genetic engineering protocols</span>  
-
      <p class="para">With our team’s biomedical background in mind, an anti-fouling chemical layer for use in the human body was chosen to test the ‘plug and play’ system. An anti-fouling hydrogel has to have little to no interaction with the human immune system, thus preventing immune responses caused by the presence of bacteria. Dibenzocyclooctyne Polyethylene glycol 10kDa (DBCO-PEG 10kDa) was chosen as the molecule to click onto OmpX to form the hydrogel because it has good anti-fouling properties and the modular length allows for easy testing on a smaller scale.</p>
+
<br>  <p class="para">The following protocols are used in the Biolab during the modification of bacteria.
-
+
 
-
      </div>
+
<br><br>
 +
 
 +
  </p>
 +
      <p class="para"><a target="_blank" href="https://static.igem.org/mediawiki/2014/6/6e/Protocol_-_Colony_PCR.pdf">Colony PCR</a>
 +
 +
<a target="_blank" href="https://static.igem.org/mediawiki/2014/b/b1/Protocol_-_FACS_%28Antibody_Titration%29.pdf">Antibody Titration with FACS</a>
 +
<br><br>
 +
 
 +
<a target="_blank"  href="https://static.igem.org/mediawiki/2014/7/7e/Protocol_-_FACS_%28DBCO-56-TAMRA%29.pdf">FACS for sorting with DBCO-TAMRA</a>
 +
<br><br>
 +
 
 +
<a target="_blank" href="https://static.igem.org/mediawiki/2014/f/f0/Protocol_-_FACS_%28DBCO-PEG_10_kDa%29.pdf">FACS for sorting with DBCO-PEG(10kDa)</a>
 +
<br><br>
 +
 
 +
<a target="_blank" href="https://static.igem.org/mediawiki/2014/a/ae/Protocol_-_Insert_%2B_Vector_Ligation.pdf">Vector Ligation</a></p>
 +
 
 +
<a target="_blank" href="https://static.igem.org/mediawiki/2014/8/81/Protocol_-_Overhang_PCR.pdf">Overhang PCR</a></p>
 +
 
 +
<a target="_blank"  href="https://static.igem.org/mediawiki/2014/0/0a/Protocol_-_PCR_Purification_of_Insert_Fragment.pdf">PCR purification of DNA fragments</a></p>
 +
 
 +
<a target="_blank" href="https://static.igem.org/mediawiki/2014/2/22/Protocol_-_Plasmid_Amplification.pdf">Plasmid Amplification</a></p>
 +
<a target="_blank" href="
 +
https://static.igem.org/mediawiki/2014/0/07/Protocol_-_Plasmid_and_gene_digestion.pdf">Plasmid Gene Digestion</a></p>
 +
 
 +
<a target="_blank" href="
 +
https://static.igem.org/mediawiki/2014/2/29/Protocol_-_Plasmid_purification.pdf">Plasmid Purification</a></p>
 +
 
 +
<a target="_blank" href="
 +
https://static.igem.org/mediawiki/2014/1/13/Protocol_-_Preparative_steps.pdf">Preparative Steps</a></p>
 +
 
 +
<a target="_blank" href="
 +
https://static.igem.org/mediawiki/2014/f/f8/Protocol_-_Protein_Expression_Curve.pdf">Protein Expression Curve</a></p>
 +
 
 +
<a target="_blank" href="
 +
https://static.igem.org/mediawiki/2014/4/45/Protocol_-_Protein_expression.pdf">Protein Expression</a></p>
 +
<a target="_blank" href="
 +
https://static.igem.org/mediawiki/2014/6/62/Protocol_-_Site_Directed_Mutagenesis.pdf">Site Directed Mutagenesis</a></p>
 +
 
 +
<a target="_blank" href="
 +
https://static.igem.org/mediawiki/2014/b/b5/Protocol_-_Transformation.pdf">Vector Transformation</a></p>
 +
 
 +
 
 +
 
 +
</div>
 +
 
      <div class="clear"></div>
      <div class="clear"></div>
      </div>
      </div>
-
<div class="cont-grid_left-img img_style">
+
     
-
    <img src="https://static.igem.org/mediawiki/2014/2/20/Igemeindhovenuitlegconceptehvresized.jpg" alt="" width="400px">
+
-
    </div>      
+
      <div class="clear"></div>
      <div class="clear"></div>
 +
     
 +
 
 +
     
<br><br>
<br><br>
 +
<div class="clear"></div>
 +
<div class="cont-grid_left">
 +
      <div class="Jasper">
 +
  <span>chemistry protocols</span>   
 +
      <p class="para">
 +
<a href="
 +
https://static.igem.org/mediawiki/2014/3/3d/Protocol_-_Monitoring_SPAAC_with_UVVIS.pdf">SPAAC reaction monitoring with UV-Visible Spectroscopy</a></p>
 +
 +
 +
</div>
 +
      <div class="clear"></div>
 +
      </div>
 +
     
 +
      <div class="clear"></div>
 +
     
 +
 
 +
     
 +
 +
<br><br>
<div class="clear"></div>
<div class="clear"></div>
<div class="cont-grid_left">
<div class="cont-grid_left">
      <div class="Jasper">
      <div class="Jasper">
-
  <span>Microfluidics</span>   
+
  <span>microfluidics protocols</span>   
-
      <p class="para">In order to precisely control the hydrogel formation, microfluidic devices are used in which the conditions are optimal to form individually encapsulated cells. This way clustering of cells is prevented and the end product will be usable beads instead of large aggregated blobs.  </p>
+
      <p class="para">Here will follow all microfluidic engineering protocols</p>
-
+
</div>
-
      </div>
+
      <div class="clear"></div>
      <div class="clear"></div>
      </div>
      </div>
-
<div class="cont-grid_left-img img_style">
+
     
-
    <img src="https://static.igem.org/mediawiki/2014/5/52/Igemeindhovenuitleg1resized.jpg" alt="" width="400px">
+
-
    </div>      
+
      <div class="clear"></div>
      <div class="clear"></div>
 +
     
 +
 
 +
     
-
 
+
<br><br>
-
 
+
-
 
+
</div>
</div>
</div>
</div>
 +
</html>
</html>
{{:Team:TU_Eindhoven/Template:Footer}}
{{:Team:TU_Eindhoven/Template:Footer}}

Latest revision as of 19:19, 31 August 2014

Project Description

Protocols

Introduction

For this year's iGEM competition numerous protocols were devoloped to guide our experiments and keep our documentation neat and tidy. Because these protocols can also be useful to other projects, we decided to publish them on our wiki. You can find information and download links on this page down below.



genetic engineering protocols

The following protocols are used in the Biolab during the modification of bacteria.

Colony PCR Antibody Titration with FACS

FACS for sorting with DBCO-TAMRA

FACS for sorting with DBCO-PEG(10kDa)

Vector Ligation

Overhang PCR

PCR purification of DNA fragments

Plasmid Amplification

Plasmid Gene Digestion

Plasmid Purification

Preparative Steps

Protein Expression Curve

Protein Expression

Site Directed Mutagenesis

Vector Transformation



chemistry protocols

SPAAC reaction monitoring with UV-Visible Spectroscopy



microfluidics protocols

Here will follow all microfluidic engineering protocols



Created by iGEM Team TU Eindhoven 2014

Retrieved from "http://2014.igem.org/Team:TU_Eindhoven/Project/protocols"