Team:Waterloo/Lab Book

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       <h2>CRISPRi</h2>
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       <img src="https://static.igem.org/mediawiki/2014/e/e2/CRISPRi_1.png"></a><br>
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    <strong>June 6, 2014</strong><br>Digested pSB1C3 backbone with EcoRI and PstI and gel purified<br>
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      <img src="https://static.igem.org/mediawiki/2014/5/53/CRISPRi_2.png"></a><br>
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    <strong>June 9, 2014</strong><br>Received synthesized sgRNA 1 and sgRNA 2 constructs from BioBasic and transformed the DNA into E. coli DH5α <br>
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      <img src="https://static.igem.org/mediawiki/2014/d/d6/CRISPR_3.png"></a><br>
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    <strong>June 11, 2014</strong><br>Inoculated sgRNA 1 and 2 colonies from the transformation into over night liquid culture for frozen stocks and miniprep<br>
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      <img src="https://static.igem.org/mediawiki/2014/f/f9/CRISPR_4.png"></a><br>
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    <strong>June 18, 2014</strong><br>Digested sgRNA 1 and 2 constructs with EcoRI and PstI, then gel purified the inserts for cloning into pSB1A3
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      <img src="https://static.igem.org/mediawiki/2014/1/1d/CRISPR_5.png"></a><br>
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Also digested sgRNA 1 with EcoRI and SpeI, and sgRNA 2 with XbaI and PstI for modified 3A assembly into pSB1A3<br>
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      <img src="https://static.igem.org/mediawiki/2014/5/5c/CRISPRi_6.png"></a><br>
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    <strong>June 19, 2014</strong><br>Digested pSB1A3 linearized backbone with EcoRI and PstI, then column purified. <br>Ligated pSB1A3 with the sgRNA 1 and 2 (E/P) inserts individually and with both sgRNA 1 (E/S) and sgRNA 2 (X/P) together <br>Ligated pSB1C3 backbone with sgRNA 1 (E/P) and sgRNA 2 (E/P) individually for BioBricks<br>
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       <img src="https://static.igem.org/mediawiki/2014/8/82/CIRSPRi_7.png"></a><br>
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    <strong>June 20, 2014</strong><br>Transformed all ligations into E. coli DH5α<br>
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      <img src="https://static.igem.org/mediawiki/2014/0/0b/CRISPRi_8.png"></a><br>
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    <strong>June 21, 2014</strong><br>Streak purified 4 colonies from each ligation<br>
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      <img src="https://static.igem.org/mediawiki/2014/3/37/CRISPRi_9.png"></a><br>
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    <strong>June 22, 2014</strong><br>Inoculated 4 colonies of each ligation into overnight liquid culture for miniprep (patched colonies onto agar plates before inoculating<br>
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    <strong>June 23, 2014</strong><br>Miniprepped and did a diagnostic digest to see which clones contained the appropriate inserts:
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sgRNA 1 in pSB1A3                          1 successful<br>
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sgRNA 2 in pSB1A3                          3 successful<br>
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sgRNA 1 and sgRNA 2 in pSB1A3       0 successful<br>
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sgRNA 1 in pSB1C3                          1 successful<br>
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sgRNA 2 in pSB1C3                          2 successful<br>
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    <strong>June 24, 2014</strong><br>WRITE HERE<br>
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    <strong>June 25, 2014</strong><br>WRITE HERE<br>
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    <strong>June 26, 2014</strong><br>WRITE HERE<br>
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    <strong>July 3, 2014</strong><br>WRITE HERE<br>
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    <strong>July 4, 2014</strong><br>WRITE HERE<br>
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    <strong>July 9, 2014</strong><br>WRITE HERE<br>
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    <strong>June 25, 2014</strong><br>WRITE HERE<br>
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      <h2>RNAi</h2>
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       <h2>YFP</h2>
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       <img src="https://static.igem.org/mediawiki/2014/1/1d/YFP_1.png"></a><br>
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      <img src="https://static.igem.org/mediawiki/2014/5/59/YFP_2.png"></a><br>
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       <h2>CONJUGATION</h2>
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       <img src="https://static.igem.org/mediawiki/2014/2/28/Conjugation_1.png"></a><br>
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       <h2>OTHER</h2>
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       <img src="https://static.igem.org/mediawiki/2014/8/8e/Other_1.png"></a><br>
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Latest revision as of 03:28, 18 October 2014

Lab Book



















Retrieved from "http://2014.igem.org/Team:Waterloo/Lab_Book"