Team:Yale/Our Project/Notebook/Week 2

From 2010.igem.org

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*This plan was eventually altered, as promoter choices were altered and no light-inducible promoter was found. <br/>
*This plan was eventually altered, as promoter choices were altered and no light-inducible promoter was found. <br/>
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To make these plasmids, we will rely on the standard iGEM assembly protocol involving restriction enzymes EcoRI, XbaI, PstI, and SpeI shown in the diagram at right, but in the first ligation the B0015 terminator will take the place of C0010 and the phsABC  gene in pSB74 will take the place of B0034. <br/>
 
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<img src="https://static.igem.org/mediawiki/2010/7/71/Yale-biobrick.jpg" />
<img src="https://static.igem.org/mediawiki/2010/7/71/Yale-biobrick.jpg" />
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To make these plasmids, we will rely on the standard iGEM assembly protocol involving restriction enzymes EcoRI, XbaI, PstI, and SpeI shown in the diagram at right, but in the first ligation the B0015 terminator will take the place of C0010 and the phsABC  gene in pSB74 will take the place of B0034. <br/>
<b> Steps to make these plasmids </b> <br/>
<b> Steps to make these plasmids </b> <br/>

Revision as of 05:54, 27 October 2010

iGEM Yale

lab notebook: week 2 (6/14-6/20)

  • Monday 6/14--Redo of 6/11 assay of bacterial growth within a narrow copper(II) concentrations after analysis of data showed uniformly poor growth.
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  • Tuesday 6/15--Results and analysis of the 6/14 narrow concentration range growth assay as well as planning for the creation of a standard iGEM plasmid bearing the thiosulfate reductase operon (phsABC).
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  • Wednesday 6/16--Checked on spotted cell survival assay, collected MOPS minimal media materials & started making component solutions
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  • Thursday 6/17--more minimal media work and meeting, started BL21 culture
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  • Friday 6/18--Arrival of plasmid pSB74, transformation of Biobricks, & copper growth assays for BL21 strain.
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  • Saturday 6/19--Redo of Biobrick transformations & analysis of BL21 copper growth assay
  • See more/less
  • Sunday 6/20--Observed growth of all transformants from 6/19, so used them and the culture containing pSB74 to inoculate 5 mL liquid cultures in LB with ampicillin. Left to grow overnight on shaker at 37˚C for miniprep the following morning.
    The activities of this day are also recorded on page 16 of the hard copy lab notebook