Difference between revisions of "Team:Waterloo/Parts"

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     <h1> Part Documentation</h1>
 
     <h1> Part Documentation</h1>
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<h2> Characterized Parts </h2>
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The xylose inducible promoter is one of the parts our team characterized this year. The reason we want to characterize this promoter is because it has been mutated via site directed mutagenesis in order to remove illegal sites. Now that the promoter has been mutated, it must be tested to see if it is expressed efficiently in the presence of xylose. The parts we are characterizing are BBa_K1323014 and BBa_K1323002.
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To characterize this variant of the xylose inducible promoter, it was cloned in front of GFP so a simple fluorescence assay could be performed. The new mutated xylose promoter should be activated in the presence of media containing 2% xylose, assuming its function is no different than the original <cite ref="Forsyth2002"></cite>. The positive control was a plasmid containing the constitutive J23101 promoter in front of GFP. The negative control in this assay was using a plasmid including GFP with no promoter. All plasmids were transformed into the DH5alpha Escherichia coli strain.
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Strains with the plasmids were grown to an OD600=0.2 and then were diluted into media and put into a plate reader. Measurements of OD600 and GFP expression were taken at intervals of 10 minutes until the positive control expressed the GFP promoter.
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<h2> References </h2>
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         <li><a href="/Team:Waterloo/Part_Collection">Part Collection</a></li>
 
         <li><a href="/Team:Waterloo/Part_Collection">Part Collection</a></li>

Revision as of 01:12, 19 September 2015

Part Documentation

Characterized Parts

The xylose inducible promoter is one of the parts our team characterized this year. The reason we want to characterize this promoter is because it has been mutated via site directed mutagenesis in order to remove illegal sites. Now that the promoter has been mutated, it must be tested to see if it is expressed efficiently in the presence of xylose. The parts we are characterizing are BBa_K1323014 and BBa_K1323002.

To characterize this variant of the xylose inducible promoter, it was cloned in front of GFP so a simple fluorescence assay could be performed. The new mutated xylose promoter should be activated in the presence of media containing 2% xylose, assuming its function is no different than the original . The positive control was a plasmid containing the constitutive J23101 promoter in front of GFP. The negative control in this assay was using a plasmid including GFP with no promoter. All plasmids were transformed into the DH5alpha Escherichia coli strain.

Strains with the plasmids were grown to an OD600=0.2 and then were diluted into media and put into a plate reader. Measurements of OD600 and GFP expression were taken at intervals of 10 minutes until the positive control expressed the GFP promoter.

References

Part Table 

       <groupparts>iGEM015_Waterloo</groupparts>

Each team will make new parts during iGEM and will submit them to the Registry of Standard Biological Parts. The iGEM software provides an easy way to present the parts your team has created. The <groupparts> tag (see below) will generate a table with all of the parts that your team adds to your team sandbox.

Remember that the goal of proper part documentation is to describe and define a part, so that it can be used without needing to refer to the primary literature. Registry users in future years should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for users who wish to know more.

Note

Note that parts must be documented on the Registry. This page serves to showcase the parts you have made. Future teams and other users and are much more likely to find parts by looking in the Registry than by looking at your team wiki.

Adding parts to the registry

You can add parts to the Registry at our Add a Part to the Registry link.

We encourage teams to start completing documentation for their parts on the Registry as soon as you have it available. The sooner you put up your parts, the better you will remember all the details about your parts. Remember, you don't need to send us the DNA sample before you create an entry for a part on the Registry. (However, you do need to send us the DNA sample before the Jamboree. If you don't send us a DNA sample of a part, that part will not be eligible for awards and medal criteria.)

What information do I need to start putting my parts on the Registry?

The information needed to initially create a part on the Registry is:

  • Part Name
  • Part type
  • Creator
  • Sequence
  • Short Description (60 characters on what the DNA does)
  • Long Description (Longer description of what the DNA does)
  • Design considerations

We encourage you to put up much more information as you gather it over the summer. If you have images, plots, characterization data and other information, please also put it up on the part page.

Inspiration

We have a created a collection of well documented parts that can help you get started.

You can also take a look at how other teams have documented their parts in their wiki:

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