From 2011.igem.org
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- | {{Team:USTC-China/temp}}
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- | {{Team:USTC-China/temp/bar4}}
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- | *Aptamer-cheZ Device Construction
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- | <p>The cheZ gene was cloned from E.coli TOP10 strain using PCR. A cassete containing a theophylline-sensitive aptamer, and the cheZ gene was assembled using PCR and was subcloned into the SpeI and PstI sites of pSB1A2 containing the lac promoter.</p>
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- | <p>Clone Primer:</p>
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- | Forward:5'-GTTTCGAATTCGCGGCCGCTTCTAGATGCAACCATCAATCAAACC-3'
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- | Reverse:5'-GTTTCCTGCAGCGGCCGCTACTAGTATTATTAAAATCCAAGTCTATCCAACAAATCGT-3'
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- | <p>Assemble Primer:</p>
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- | <p>Forward:5'-GTTTCGAATTCGCGGCCGCTTCTAGGGTGATACCAGCATCGTCTTGATGCCCTTGGCAGCACCCCGCTGCAAGACAACAAG ATGCAACCATCAATCAAACC-3'</p>
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- | <p>Reverse:5'-GTTTCCTGCAGCGGCCGCTACTAGTATTATTAAAATCCAAGACTATCCAACAAATCGT-3'</p>
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- | <p>Assemble condition: 96℃ 10min, (96℃ 30s, 56℃ 30s, 72℃ 1min)16 cycles and each cycle the annealing temperature increase 1℃, (96℃ 30s, 72℃ 30s, 72℃ 1min)20 cycles,72℃ 10min and holding at 4℃.</p>
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Latest revision as of 18:08, 5 October 2011