Team:UGent/Results
From 2013.igem.org
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We constructed plasmid pSB6A1-T7<i>ccdB</i> for use in CIChE. Through this plasmid, pressure can be put on the CIChE strains simply by adding IPTG. | We constructed plasmid pSB6A1-T7<i>ccdB</i> for use in CIChE. Through this plasmid, pressure can be put on the CIChE strains simply by adding IPTG. | ||
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We also purified plasmids p5SpFRT-T7<i>ccdB</i>, p10SpFRT-T7<i>ccdB</i>, p20SpFRT-T7<i>ccdB</i> from strains we obtained from Inbio. These plasmids have different copy numbers and they too can be used in chromosomal evolution. | We also purified plasmids p5SpFRT-T7<i>ccdB</i>, p10SpFRT-T7<i>ccdB</i>, p20SpFRT-T7<i>ccdB</i> from strains we obtained from Inbio. These plasmids have different copy numbers and they too can be used in chromosomal evolution. | ||
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<h1> Strains with <i>ccdA-gfp</i> construct </h1> | <h1> Strains with <i>ccdA-gfp</i> construct </h1> | ||
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- | The following strain containing the construct to be duplicated (<i>ccdA-gfp</i>) was constructed (KI experiment 1):</p> | + | The following strain containing the construct to be duplicated (<i>ccdA-gfp</i>) was constructed (KI experiment 1): |
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+ | <table> | ||
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+ | <li> Colony PCR on 48 colonies using crimson taq polymerase with two primer pairs: MDM0141/MDM0010 and MDM0046/MDM123. Expected fragments: 550bp & 2450 bp. Analytic gel: <b> 4 positives </b> | ||
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+ | <img src="https://static.igem.org/mediawiki/2013/0/02/UGent_2013_CPCR_KI.jpg" width="300"/></a> | ||
+ | </td> | ||
+ | </tr> | ||
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+ | <li> Colony PCR on 4 positive and 4 negative colonies using Emerald polymerase with out primers: MDM0046/MDM0010. Expected fragment: ca. 5500 bp. Analytic gel: <b> 1 positive: colonie 35 </b>.</li></ul> | ||
+ | </td> | ||
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+ | <img src="https://static.igem.org/mediawiki/2013/3/35/UGent_2013_CPCR_KI_out.png" width="200"/></a></li> | ||
+ | </td> | ||
+ | </tr> | ||
+ | </table> | ||
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<p><center><i>E. coli</i> MG1655 ΔendA DE3 ccdA-Pmb1GFP-CmFRT (called strain 8)</center></p> <p> | <p><center><i>E. coli</i> MG1655 ΔendA DE3 ccdA-Pmb1GFP-CmFRT (called strain 8)</center></p> <p> | ||
We have two versions of this strain: 8 and MDM.Cm. The two strains were used in our further experiments as to test possible differences between the two versions. | We have two versions of this strain: 8 and MDM.Cm. The two strains were used in our further experiments as to test possible differences between the two versions. | ||
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</p> | </p> | ||
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<h1> CIChE Strains</h1> | <h1> CIChE Strains</h1> | ||
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Revision as of 21:08, 4 October 2013
Plasmids containing T7-ccdB
We constructed plasmid pSB6A1-T7ccdB for use in CIChE. Through this plasmid, pressure can be put on the CIChE strains simply by adding IPTG.
Strains with ccdA-gfp constructThe following strain containing the construct to be duplicated (ccdA-gfp) was constructed (KI experiment 1):
We have two versions of this strain: 8 and MDM.Cm. The two strains were used in our further experiments as to test possible differences between the two versions.
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