Difference between revisions of "Team:NYU Shanghai/Protocols"

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     <h6><font color="#d66">Building our Construct</font></h6>
 
     <h6><font color="#d66">Building our Construct</font></h6>
     <h6>Note: If using construct with araC/pBAD promoter, DO NOT USE SOC MEDIA. Glucose inhibits the uptake of arabinose, and will inhibit promoter induction.</h6>
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     <p>Note: If using construct with araC/pBAD promoter, DO NOT USE SOC MEDIA. Glucose inhibits the uptake of arabinose, and will inhibit promoter induction.</p>
 
     <br><img src="https://static.igem.org/mediawiki/2015/d/df/NYU_Shanghai_Chromo_Procedure.png" width="800">
 
     <br><img src="https://static.igem.org/mediawiki/2015/d/df/NYU_Shanghai_Chromo_Procedure.png" width="800">
 
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Revision as of 16:14, 18 September 2015

Protocols

We built our constructs from pre-made biobrick parts. Our overall conclusion is that 3A assembly is generally inefficient, and an insufficient method for adding small parts (such as a terminator) to a larger construction within pSB1C3. We learned that ratios were extremely important in the process of 3A Assembly, and we made a summary sheet of the equations we used in pre-digest and pre-ligation that accounts for digest dilution and amount needed to ensure results are seen on a gel, not just ligation ratios. We wished we used gibson assembly.


Making Color

Recipes

3A Assembly

Calculations (pdf)