Wiki/Team:Warsaw/primers

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<h1>Primers</h1>
<h1>Primers</h1>
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<h2>'One-armed bandit' strain preparation</h2>
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<h2>cloning primers</h2>
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<h4>AID and T7 cloning</h4>
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<h4>cloning primers</h4>
<pre>
<pre>
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<a name="T7lLinkB">T7lLinkB</a>  5' CGGATCCGGCGGTGGCGGTAGTGGAGGAGGTGGGTCTGGAGGCGGAGGGAACACGATTAACATCGCTAAGAAC 3'
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<a name="croboxF">croboxF</a>  5' ATCTAGATACCTCTGGCGGTGATACTAGTGT 3'
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<a name="T7lRBSHi">T7lRBSHi</a>  5' CACAAGCTTAGGAGGTGTACTAGATGAACACGATTAACATCGCTAAG 3'
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<a name="croboxR">croboxR</a>  5' ACACTAGTATCACCGCCAGAGGTATCTAGAT 3'
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<a name="T7pXbSal">T7pXbSal</a>   5' TCTGTCGACTCTAGATTACGCGAACGCGAAGTCC 3'
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<a name="invF">invF</a>     5' ATCTAGATTATATTGACAGCGCACAGAGCGGGAA 3'
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<a name="AIDlNrH">AIDlNrH</a>   5' CGTAAGCTTAGGAGGTGTAGACCATGGACAGCCTCTTGATG 3'
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<a name="invR">invR</a>     5' GTACTAGTCATGGCTGGGGCTTCAGAAAAATATGAT 3'
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<a name="AIDpLinB">AIDpLinB</a>   5' CCACTACCGCCACCGCCGGATCCGCCACCTCCGCTGCCTCCACCACCAAGTCCCAAAGTACGAAATGC 3'
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<a name="hlyF">hlyF</a>     5' TGTCTAGAATGAAAAAAATAATGCTAGTT  3'
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<a name="hlyR">hlyR</a>      5' GACTAGTTTATTCGATTGGATTATCTA  3'
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<a name="croF">croF</a>      5' ATCTAGAATGGAACAACGCATAACC  3'
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<a name="croR">croR</a>      5' GTACTAGTGGGCTGGAATGTGTAAG  3'
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<a name="phoF">phoF</a>      5' ATCTAGAATGATGCGCGTACTGGTTGTAG  3'
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<a name="phoR">phoR</a>      5' TTACTAGTGATGCGTGGAAGAACGCACAG  3'
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<a name="mgtCF">mgtCF</a>    5' ATCTAGATCCATTTTTTCTGGAAGAATAAG  3'
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<a name="mgtCR">mgtCR</a>    5' GTACTAGTCCTCGTCATGTTGTTTTATTTTTT 3'
</pre>
</pre>
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<h4>Sequencing of pMPM-T5</h4>
 
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<pre>
 
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<a name="pMPMT5seqF">pMPMT5seqF</a>  5' CTACTGTTTCTCCATACCCG  3'
 
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<a name="pMPMT5seqR">pMPMT5seqR</a>  5' GAAAATCTTCTCTCATCCGC 3'
 
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<a name="AIDseqP">AIDseqP</a>    5' GCTTCGGCGCATCCTTTTG 3'
 
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</pre>
 
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<h4>Sequencing of β-galactosidase from pZC320</h4>
 
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<pre>
 
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<a name="pZCseqL">pZCseqL</a>    5' CTTGACCCGCAGTTGCAAAC 3'
 
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<a name="pZCseqR">pZCseqR</a>    5' CCCAACTGATCTTCAGCATC 3'
 
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</pre>
 
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<h4>Sequencing of the T7 polymerase gene</h4>
 
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<pre>
 
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<a name="T7-2seq">T7-2seq</a>  5' AGGTGCGCTGGCTGGCATC 3'
 
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<a name="T7-3seq">T7-3seq</a>  5' AAGGAAGGTTACTACTGGCT 3'
 
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<a name="T7-4seq">T7-4seq</a>  5' CAAAGAGTTCGGCTTCCGTC 3'
 
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<a name="T7Rseq">T7Rseq</a>    5' GCTTCAACCGCAGCTACCAC 3'
 
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</pre>
 
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<h2>'Hunter and prey' strain preparation</h2>
 
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<h4>Fusion of protein A and ompA and alpha or omega part of beta-lactamase</h4>
 
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<pre>
 
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<a name="AlphaL_link">alphaL_link</a>  5' CAGTGGTGGAAGCGGTGGCGGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGTCACCCAGAAACGCTGGTGAAAG 3'
 
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<a name="AlphaP_XB">alphaP_XB</a>    5' GG GGATCCGGTCTAGATTATTCGCCAGTTAATAGTTTGCG 3'
 
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<a name="OmegaL_link">omegaL_link</a>  5' CAGTGGTGGAAGCGGTGGCGGAGGATCTGGAGGAGGTGGTTCTGGAGGTGGAGGTCTACTTACTCTAGCTTCCCGG 3'
 
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<a name="OmegaP_EPB">omegaP_EPB</a>  5' AAGGATCCAACTGCAGAAGAATTCTTACCAATGCTTAATCAGTG 3'
 
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<a name="OmpaL_N">ompaL_N</a>      5' CATATGAAAGCTACTAAACTGGTACTGGG 3'
 
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<a name="OmpaP_link">ompaP_link</a>  5' GCCACCGCTTCCACCACTGCGGCCGCCAGAGCTCCCTCCTCCAGAACCTCCGCCGTTGTCCGGACGAGTGCCGATG 3'
 
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</pre>
 
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<h4>Cloning of A and/or Z protein to expression vector (pET15b/pET24a)</h4>
 
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<pre>
 
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<a name="AlphaP+link+Sac">AlphaP+link+Sac</a>  5' GG GAGCTC CC ACCACCCCCACCCCCGCTACCTC  3'
 
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<a name="AlphaL+Nde">AlphaL+Nde</a>        5' CGGG CATATG CACCCAGAAACGCTGGTGAAAG  3'
 
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</pre>
 
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<h4>Mutagenesis of the A protein</h4>
 
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<pre>
 
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<a name="ADelL+KpnI">ADelL+KpnI</a>  5' AGGTACCCATGTAAGATCTCATAGAACGCG  3'
 
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<a name="ADelP">AdelP</a>      5' AACTTAAACGAAGAACAACGAAACGCC  3'
 
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<a name="AmutL+KpnI">AmutL+KpnI</a>  5' TCGTTTAAGTTAGGTACCGGTGAGATCTCAGAGGACGCGTTTTGTTGTTCTTTGTTG  3'
 
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<a name="AmutP">AmutP</a>      5' AGAACAAGGAAACGCCTTCGCCCAAAGTTTAGCAGATGACCCAAGCCAAAGCGC  3'
 
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</pre>
 
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<h4>Fusion of the A protein with the OmpaA protein and the alpha/omega part of beta-lactamase protein</h4>
 
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<pre>
 
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<a name="AL+link10+homo2">AL+link10+homo2</a>    5' TCTGGAGGTGGAGGTAGCGGGGGTGGGGGTGGTTCGGGTGGAGGTGGTAAAACCGCGGCTCTTGCGC  3'
 
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<a name="AlphaP+link10+homo2">AlphaP+link10+homo2</a> 5' CCGCTACCTCCACCTCCAGAACCACCTCCTCCGCTTCCTCCGCCACCTTCGCCAGTTAATAGTTTGCG  3'
 
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<a name="OmegaP+link10+homo2">OmegaP+link10+homo2</a> 5' CCGCTACCTCCACCTCCAGAACCACCTCCTCCGCTTCCTCCGCCACCCCAATGCTTAATCAGTGAGGC  3'
 
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<a name="AL+SacI">AL+SacI</a>            5' GG GAGCTCTAAAACCGCGGCTCTTGCGCAAC  3'
 
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<a name="AP+NotI">AP+NotI</a>            5' AA GCGGCCGCCGGTTGACTTCCCCGCGGAATTC  3'
 
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<a name="AlphaL+SacI">AlphaL+SacI</a>        5' GG GAGCTCTCACCCAGAAACGCTGGTGAAAG  3'
 
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<a name="OmegaL+SacI">OmegaL+SacI</a>        5' GGGAGCTCGCTACTTACTCTAGCTTCCCGG  3'
 
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<a name="AlphaPlinkSac">AlphaPlinkSac</a>      5' TTGAGCTCCCTCCACCTCCGCTACCTCCACCAGAACCACCTCCTCCGCTTCCGGTACCTTCGCCAGTTAATAGTTTGC  3'
 
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</pre>
 
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<h2>Preparation of the BioBricks Standard Parts</h2>
 
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<pre>
 
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<a name="AIDpSNP">AIDP_SpeNotPst</a>      5' AAACTGCAGCGGCCGCTACTAGTATCAAAGTCCCAAAGTACGAAATG  3'
 
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<a name="AL_BNXNE">AL_BNXNE</a>            5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGGGATCCAAAACCGCGGCTCTTGCGCAAC  3'
 
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<a name="AlphaPSpe">AlphaPSpe</a>            5' GCACTAGTATTATTCGCCAGTTAATAGTTTGCG  3'
 
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<a name="APSacSpe">APSacSpe</a>            5' GGACTAGTAGAGCTCCCGTCTACTTTCGGCGCCTGAGC  3'
 
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<a name="LinLSXNE">LinLSXNE</a>            5' ATGAATTCGCGGCCGCTTCTAGAGGGAGCTCTGGTGGAAGCGGTGGCGGAGGATC  3'
 
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<a name="LinP_BS">LinP_BS</a>              5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC  3'
 
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<a name="LinP_BS2">LinP_BS2</a>            5' ATACTAGTACCGGATCCAGAACCTCCTCCACCGCTACCTCCACCTCCAGAAC  3'
 
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<a name="LinPBSNP">LinPBSNP</a>            5' AGCCTGCAGCGGCCGCTACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTACC  3'
 
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<a name="OmegPSpe">OmegPSpe</a>            5' GCACTAGTATTACCAATGCTTAATCAGTGAG  3'
 
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<a name="OmegLNde">OmegLNde</a>            5' CAGCCATATGCTACTTACTCTAGCTTCCCGG  3'
 
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<a name="OmpLNXNE">OmpLNXNE</a>            5' TAGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3'
 
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<a name="OmLNXNEB">OmLNXNEB</a>            5' ATTAGATCTGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTAC  3'
 
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<a name="PETt7L_XNE">pETt7L_XNE</a>          5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAATTAATACG  3'
 
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<a name="PlacL_XNE">PlacL_XNE</a>            5' TAGAATTCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCC  3'
 
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<a name="AP+XhoI+SpeI">AP+0XhoI+SpeI</a>        5' AAACTAGTCTCGAGGGTTGACTTCCCCGCGGAATTC  3'
 
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<a name=AIDl>AIDl</a>                5' TATCTAGAACCATGGACAGCCTCTTG 3'
 
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<a name=AraCl>AraCl</a>                5' GCTCTAGATTATGACAACTTGACGGC 3'
 
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</pre>
 
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Latest revision as of 10:30, 18 July 2009


Return to notebookReturn to the previous page

Primers

cloning primers

cloning primers

croboxF   5' ATCTAGATACCTCTGGCGGTGATACTAGTGT  3'
croboxR   5' ACACTAGTATCACCGCCAGAGGTATCTAGAT  3'
invF      5' ATCTAGATTATATTGACAGCGCACAGAGCGGGAA  3'
invR      5' GTACTAGTCATGGCTGGGGCTTCAGAAAAATATGAT  3'
hlyF      5' TGTCTAGAATGAAAAAAATAATGCTAGTT  3'
hlyR      5' GACTAGTTTATTCGATTGGATTATCTA  3'
croF      5' ATCTAGAATGGAACAACGCATAACC  3'
croR      5' GTACTAGTGGGCTGGAATGTGTAAG  3'
phoF      5' ATCTAGAATGATGCGCGTACTGGTTGTAG  3'
phoR      5' TTACTAGTGATGCGTGGAAGAACGCACAG  3'
mgtCF     5' ATCTAGATCCATTTTTTCTGGAAGAATAAG  3'
mgtCR     5' GTACTAGTCCTCGTCATGTTGTTTTATTTTTT  3'


Retrieved from "http://2009.igem.org/Wiki/Team:Warsaw/primers"