Team:NTNU Trondheim/Protocols

From 2014.igem.org

(Difference between revisions)
Line 305: Line 305:
</li>
</li>
<li class="pg-project_background">
<li class="pg-project_background">
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project">Background</a>
+
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project/Modelling">Modelling</a>
-
</li>
+
-
<li class="pg-project_wetlab">
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project">Wet Lab</a>
+
-
</li>
+
-
<li class="pg-project_drylab">
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project">Dry Lab</a>
+
</li>
</li>
<li class="pg-project_hprac">
<li class="pg-project_hprac">
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project">Human Practices</a>
+
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project/Outreach">Outreach</a>
</li>
</li>
<li class="pg-project_future">
<li class="pg-project_future">
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project">Future Applications</a>
+
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project/Future">Future Applications</a>
</li>
</li>
<li class="pg-project_safety">
<li class="pg-project_safety">
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project">Safety</a>
+
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project/Safety">Safety</a>
 +
</li>
 +
<li class="pg-project_safety">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Judging">Judging criteria</a>
</li>
</li>
</ul>
</ul>
</li>
</li>
-
<li class="has-flyout pg-outreach pg-outreach_presentations pg-outreach_panels pg-outreach_awareness">
+
<li class="pg-outreach pg-outreach_presentations pg-outreach_panels pg-outreach_awareness">
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Team">Team</a>
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Team">Team</a>
</li>
</li>
-
<li class="has-flyout pg-notebook pg-notebook_protocols pg-notebook_plasmids">
+
<li class="pg-notebook pg-notebook_protocols pg-notebook_plasmids">
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook">Notebook</a>
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook">Notebook</a>
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook" class="flyout-toggle"><span> </span></a>
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook" class="flyout-toggle"><span> </span></a>
 +
</li>
 +
<li class="pg-outreach pg-outreach_presentations pg-outreach_panels pg-outreach_awareness">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Protocols">Protocols</a>
 +
</li>
 +
<li class="has-flyout pg-outreach pg-outreach_presentations pg-outreach_panels pg-outreach_awareness">
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Sponsors">Acknowledgements</a>
 +
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Project" class="flyout-toggle"><span> </span></a>
<ul class="flyout">
<ul class="flyout">
-
<li class="pg-notebook">
+
<li class="pg-project">
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook">Overview</a>
+
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Acknowledgements/Sponsors">Sponsors</a>
</li>
</li>
-
<li class="pg-notebook_protocols">
+
<li class="pg-project_background">
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook">Protocols</a>
+
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Acknowledgements/Attributions">Attributions</a>
-
</li>
+
-
<li class="pg-notebook_plasmids">
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Notebook">Plasmids</a>
+
</li>
</li>
</ul>
</ul>
-
</li>
 
-
<li class="has-flyout pg-team pg-team_bios pg-team_sponsors pg-team_gallery pg-team_attributions">
 
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Protocols">Protocols</a>
 
-
</li>
 
-
<li class="has-flyout pg-team pg-team_bios pg-team_sponsors pg-team_gallery pg-team_attributions">
 
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim/Sponsors">Sponsors</a>
 
</li>
</li>
<li class="pg-badge">
<li class="pg-badge">
Line 357: Line 352:
</li>
</li>
-
<!--li class="has-flyout">
+
-
<a style="visibility:none"></a>
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim">
+
-
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim"><span> </span></a>
+
-
<ul class="flyout right" style="display:none;background:none;border:none;-webkit-box-shadow:none;-moz-box-shadow:none;box-shadow:none">
+
-
<li class="pg-badge">
+
-
<div class="badge-wrapper">
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim">
+
-
<img style="margin-left:7px;margin-top:2px;height:56px;" src="https://static.igem.org/mediawiki/2013/1/1c/CornellOrganofoamBadge.png">
+
-
</a>
+
-
</div>
+
-
<a href="https://2014.igem.org/Team:NTNU_Trondheim" style="margin-top:-50px">2013: Organofoam</a>
+
-
</li>
+
-
<li class="pg-badge_home">
+
-
<div class="badge-wrapper">
+
-
<a href="http://igem.engineering.cornell.edu">
+
-
<img style="margin-left: 2px;margin-top: 2px;height: 56px;" src="http://igem.engineering.cornell.edu/img/CornellBadgeLogo2.png">
+
-
</a>
+
-
</div>
+
-
<a href="http://igem.engineering.cornell.edu">Team Website</a>
+
-
</li>
+
-
<li class="pg-badge_2012">
+
-
<div class="badge-wrapper">
+
-
<a href="https://2012.igem.org/Team:Cornell">
+
-
<img style="margin-left:2px;margin-top:-1px;height:65px;" src="https://static.igem.org/mediawiki/2012/7/70/Cornell2012Badge.png">
+
-
</a>
+
-
</div>
+
-
<a href="https://2012.igem.org/Team:Cornell">2012: SAFE BET</a>
+
-
</li>
+
-
<li class="pg-badge_2011">
+
-
<div class="badge-wrapper">
+
-
<a href="https://2011.igem.org/Team:Cornell">
+
-
<img style="margin-left:4px;margin-top:8px;height:44px;" src="https://static.igem.org/mediawiki/2013/4/46/CornellBiofactory.png">
+
-
</a>
+
-
</div>
+
-
<a href="https://2011.igem.org/Team:Cornell">2011: Biofactory</a>
+
-
</li>
+
-
<li class="pg-badge_2010">
+
-
<div class="badge-wrapper">
+
-
<a href="https://2010.igem.org/Team:Cornell">
+
-
<img style="margin-left:-3px;margin-top:6px;height:60px;" src="https://static.igem.org/mediawiki/2013/3/3c/CornellOmgOmvs.png">
+
-
</a>
+
-
</div>
+
-
<a href="https://2010.igem.org/Team:Cornell">2010: OMG OMVs</a>
+
-
</li>
+
-
<li class="pg-badge_2009">
+
-
<div class="badge-wrapper">
+
-
<a href="https://2009.igem.org/Team:Cornell">
+
-
<img style="margin-left:0px;height:60px;" src="https://static.igem.org/mediawiki/2013/a/a0/CornellCadmium.png">
+
-
</a>
+
-
</div>
+
-
<a href="https://2009.igem.org/Team:Cornell" style="font-size:20px !important">2009: Cadmium Sensor</a>
+
-
</li>
+
-
</ul>
+
-
</li-->
+
</ul>
</ul>
<div class="clearfix"></div>
<div class="clearfix"></div>
Line 423: Line 363:
findNames();
findNames();
loadHref();
loadHref();
-
$('li.pg-notebook\').addClass('');
+
$('li.pg-notebook').addClass('');
});
});
</script>
</script>

Revision as of 12:26, 15 October 2014

Team:NTNU Trondheim/Protocols - 2014.igem.org

 

Team:NTNU Trondheim/Protocols

From 2014.igem.org

Team:NTNU_Trondheim/Protocols - 2014.igem.org

 

Team:NTNU_Trondheim/Protocols

From 2014.igem.org

NTNU Genetically Engineered Machines

Protocols

Filter by subteam:
show all categories
show technical details

"_"
Media
Techniques
Plasmids
Organisms
Calculations
only
only
only
only
only

Liquid Broth (LB)

Recipe
Antibiotic additions
Antibiotic Stock concentration Final concentration Dillution factor Solvent Storage temperature
Ampicillin 50 mg / mL 50 μg / mL 1000 Filter sterilized H2O 4 °C
Chloramphenicol30 mg / mL30 μg / mL 1000Ethanol-20 °C
Kanamycin50 mg / mL30 μg / mL1000Filter sterilized H2O4 °C
Spectinomycin50 mg / mL50 μg / mL1000Filter sterilized H2O4 °C

Ingredients:

  • Tryptone (10g)
  • NaCl (10g)
  • Yeast Extract (5g)

  1. Fill with 1 L of distilled / filtered H2O.
  2. Autoclave at 121 °C for 20 minutes.
  3. Add antibiotics if needed, after the medium has cooled down.

Super Optimal Broth (S.O.B.)

Recipe
show technical details
{{{tech}}}

Made LB plates with ampicillin and ampicillin + kanamycin.

yB medium

Recipe
show technical details
{{{tech}}}

Ingredients:

  • Yeast extract (2.5g)
  • Bactotryptone (10g)
  • KCL (0.38g)

  1. Fill with 0.5 L of distilled / filtered H2O.
  2. Add KOH until the pH is 7.4, then autoclave at 121 °C for 20 minutes.
  3. Add 17 mL sterile 1M MgSO4

Gibson Assembly

Protocol from New England Biolabs
show technical details
{{{tech}}}


  1. Set up the following reaction on ice:

    2.   Recommended Amount of Fragments Used for Assembly
    2-3 Fragment Assembly 4-6 Fragment Assembly Positive Control**
    Total Amount of Fragments 0.02–0.5 pmols*
    X μl    		 0.2–1 pmols*
    X μl    		 10 μl
    Gibson Assembly Master Mix (2X) 10 μl 10 μl 10 μl
    Deionized H2O 10-X μl 10-X μl 0
    Total Volume 20 μL*** 20 μL*** 20 μL
    * Optimized cloning efficiency is 50–100 ng of vectors with 2–3 fold of excess inserts. Use 5 times more of inserts if size is less than 200 bps. Total volume of unpurified PCR fragments in Gibson Assembly reaction should not exceed 20%.
    ** Control reagents are provided for 5 experiments.
    *** If greater numbers of fragments are assembled, additional Gibson Assembly Master Mix may be required.


  2. Incubate samples in a thermocycler at 50°C for 15 minutes when 2 or 3 fragments are being assembled or 60 minutes when 4-6 fragments are being assembled. Following incubation, store samples on ice or at –20°C for subsequent transformation.

    Note: Extended incubation up to 60 minutes may help to improve assembly efficiency in some cases (for further details see FAQ section).

  3. Transform NEB 5-alpha Competent E. coli cells (provided with the kit) with 2 μL of the assembly reaction, following the transformation protocol.

Retrieved from "https://2014.igem.org/Team:NTNU_Trondheim/Protocols"

Retrieved from "https://2014.igem.org/Team:NTNU_Trondheim/Protocols"

Retrieved from "http://2014.igem.org/Team:NTNU_Trondheim/Protocols"