Difference between revisions of "Team:UMaryland/HokSok"
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+ | <p style="font-size:32px;text-align:center;font-family:Verdana, Geneva, sans-serif;"><b>Purpose</b></p> | ||
+ | <br> | ||
+ | <p style="font-size:24px" | ||
<p style="font-size:32px;text-align:center;font-family:Verdana, Geneva, sans-serif;"><b>Hok/Sok Construct</b></p> | <p style="font-size:32px;text-align:center;font-family:Verdana, Geneva, sans-serif;"><b>Hok/Sok Construct</b></p> | ||
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<p style="font-size:24px">Prior to experimentation, we had to insert the Hok-Sok construct into pSB1C3 in order to make it a BioBrick. We originally planned to PCR amplify the cassette out of the R1 plasmid of <i>E. coli</i>, but we were unable to find an suitable wild-type strain that was easily available. Instead, we turned to synthesizing the construct as a gBlock from IDT. As a 580 bp dsDNA fragment, it was suitable for addition via Gibson Assembly into pSB1C3. After subsequent transformation, miniprep, and confirmation sequencing, we had the first piece of our testing puzzle.</p> | <p style="font-size:24px">Prior to experimentation, we had to insert the Hok-Sok construct into pSB1C3 in order to make it a BioBrick. We originally planned to PCR amplify the cassette out of the R1 plasmid of <i>E. coli</i>, but we were unable to find an suitable wild-type strain that was easily available. Instead, we turned to synthesizing the construct as a gBlock from IDT. As a 580 bp dsDNA fragment, it was suitable for addition via Gibson Assembly into pSB1C3. After subsequent transformation, miniprep, and confirmation sequencing, we had the first piece of our testing puzzle.</p> | ||
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− | <p style="font-size:24px;text-align:center;font-family:Verdana, Geneva, sans-serif;">Section Summary | + | <p style="font-size:24px;text-align:center;font-family:Verdana, Geneva, sans-serif;">Section Summary |
<ul> | <ul> | ||
<li> 1. Full Hok-Sok sequence taken from Gerdes et al. Sequence is originally from the R1 plasmid of <i>E. coli K-12</i>.</li> | <li> 1. Full Hok-Sok sequence taken from Gerdes et al. Sequence is originally from the R1 plasmid of <i>E. coli K-12</i>.</li> | ||
<li> 2. g-Block of Hok-Sok sequence ordered from Integrated DNA Technologies, then assembled into pSB1C3 using the Gibson Assembly method.</li> | <li> 2. g-Block of Hok-Sok sequence ordered from Integrated DNA Technologies, then assembled into pSB1C3 using the Gibson Assembly method.</li> | ||
− | </ul> | + | </ul></p> |
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Revision as of 06:12, 18 September 2015