Team:St Andrews/Procedure

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        <header class="jumbotron subhead" id="overview">
 
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            <h1>Lab Book</p>
 
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<p></p>
 
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<h2>Procedure</h2>
 
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            </header>
 
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<div class="tabbable">
 
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  <ul class="nav nav-tabs">
 
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    <li class="active"><a href="#tab1" data-toggle="tab">General Methods</a></li>
 
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    <li><a href="#tab2" data-toggle="tab">ω-3 Synthesis</a></li>
 
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    <li><a href="#tab3" data-toggle="tab">Metal-binding peptides</a></li>
 
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  </ul>
 
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  <div class="tab-content">
 
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    <div class="tab-pane active" id="tab1">
 
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      <dl class="dl-horizontal"> 
 
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        <dt>date</dt> 
 
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        <dd>what we did</dd> 
 
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        <dt>date</dt> 
 
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        <dd>what we did</dd>
 
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        <dt>date</dt> 
 
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        <dd>what we did</dd> 
 
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      </dl> 
 
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      <p>anyone really keen to write this up, up to protein visualization? Don't forget to mention the differing restriction enzymes and vectors of each team. I can add in links to the protocols page.</p>
 
-
      <p>Please also refer to our <a href="https://2012.igem.org/Team:St_Andrews/Lab-book"><font color="gray">Protocols</font></a> page.</p>
 
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<BR>&nbsp;</BR>
 
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<BR>&nbsp;</BR>
 
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<BR>&nbsp;</BR>
 
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<BR>&nbsp;</BR>
 
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<BR>&nbsp;</BR>
 
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<BR>&nbsp;</BR>
 
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<BR>&nbsp;</BR>
 
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<BR>&nbsp;</BR>
 
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<BR>&nbsp;</BR>
 
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<BR>&nbsp;</BR>
 
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    </div>
 
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    <div class="tab-pane" id="tab2">
 
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<BR>&nbsp;</BR>
 
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<span class="label"><a href="#Sequences"><font color="white">Sequences</font></a></span>&nbsp;&nbsp;&nbsp;<span class="label"><a href="#Primers"><font color="white">Primers</font></a></span>&nbsp;&nbsp;&nbsp;<span class="label"><a href="#Sequencing"><font color="white">Sequencing results</font></a></span>&nbsp;&nbsp;&nbsp;<span class="label"><a href="#Lipid"><font color="white">Lipid analysis</font></a></span>
 
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<BR>&nbsp;</BR>
 
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      <p>Start from lipid extraction</p>
 
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      <p>primers, sequence results</p>
 
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<BR>&nbsp;</BR>
 
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<a name="Sequences"><span class="label">Sequences</span></a>
 
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<div class="btn-toolbar">
 
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<div class="btn-group">
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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    Δ12 T. Cruzi
 
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    <span class="caret"></span>
 
-
  </a>
 
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  <ul class="dropdown-menu">
 
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    <div class="span5"><p><font size="1">
 
-
atgtcgcgggttgacaatttaactgtcgcgccgggtccaccggatgtcatgaaggcggtgcttaagtctgggcgcaacaaggagcacaat
 
-
gtcattgtgccttcctccgcactgaccattggagaaattcaggagaaaatcccggtaaagtactttgaacgcaacactacccggagcgtc
 
-
atgtttcttttacgcgacctcgcacaggtggcgttgacgtacgccattatgtacgccgtcgccttgccacttgccacttccatggaagtttctg
 
-
cagcaagaacgttggccgacggtggggtgttatcattgatggcaggtacagccatgacgacagcggcgtggcttttaaaggcagtgttgtg
 
-
ggcggtgttttggtttgtacagggacttaacggcactgcgctttgggtcctggctcacgaatgcggccatcaggcgttcagccccatgaaag
 
-
gtttgaatgacgccgttggcatgattttgcactctgcgctgctcgtgccgtaccacagctggcgcatcacccacggcggccaccacaaac
 
-
acacgaatcacctcacgaaggatcttgtgtttgttccagacaaacgaaacgcggttgtggagctcgtggaggaggcgccgctggtgttatt
 
-
aattcaattattgctgatttttctctttggttggccggcacatcttctttttaatgcctctggacaggaatttggccgactcgcgagccactttgac
 
-
cccggcgctccatttttccgcagcgaagaccgtcacgatattgtcctgtcgaatgttgggattgtcagcgcgttatttgtcattttctccagcgt
 
-
ttaccgctttggttttacaaatgttttctgctggtacattgtaccgtacctctgggtgaacttttggcttgtgtacattacatacctgcagcacacg
 
-
gatatacgcattcctcactacacacatgagcactggacgtttgttcgcggtgcattggcggctgtggacagggactacggctttgtccttaac
 
-
acatggctccatcacatcaatgattcccacgtggtacatcacctctttagcaaaatgccacactacaacgcaatccaggtgacaagaaag
 
-
tacattcgtgagatactgggtgccacatacattacggatgagaggtcactgtggaagatgctctgggaacagcgtagagagtgccgctatg
 
-
ttgttcccgcagagggcgtctgtgtctttcatgggtaa</font></p></div>
 
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  </ul>
 
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</div>
 
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<div class="btn-group">
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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    Δ12 Synechocystis
 
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    <span class="caret"></span>
 
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  </a>
 
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  <ul class="dropdown-menu">
 
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    <div class="span5"><p><font size="1">atgactgccacgattcccccgttgacaccaacggtaacgcccagcaatcccgatcgcccg
 
-
attgcggatctcaaactacaagacatcattaaaaccctgcccaaggaatgcttcgagaaa
 
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aaagcgagcaaagcctgggcttctgttttgattaccctaggggcgatcgccgtgggctat
 
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ttgggcattatttatctgccctggtactgcttgcccattacctggatctggacagggaca
 
-
gccttaacgggggccttcgttgtcggccatgactgtggccatcgctcctttgctaaaaaa
 
-
cgctgggtcaatgatttagtgggacatatcgcttttgctcccctcatctaccctttccat
 
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agctggcgcctactccacgaccaccatcacctccacaccaacaaaattgaggttgataac
 
-
gcctgggatccctggagtgtggaagctttccaagccagcccggcgatcgtccggcttttt
 
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tatcgggccatccggggtcccttctggtggactggttccattttccattggagcttaatg
 
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cacttcaaactttccaactttgcccaaagggaccgcaataaagtcaaattatccattgcc
 
-
gttgtcttcctgtttgcggcgatcgcctttcctgccctaattatcaccacaggggtgtgg
 
-
ggtttcgtcaaattttggctaatgccctggttggtgtatcacttttggatgagcactttt
 
-
accattgtgcaccacaccattcccgaaattcgtttccgtcccgccgccgattggagtgcc
 
-
gccgaagcccagttaaatggtactgttcactgcgattatccccgttgggtggaagtgctc
 
-
tgccatgacatcaacgtccatattccccaccacctctccgttgccatcccttcctataac
 
-
ctacgactagcccacggaagtttaaaagaaaactggggaccttttctttacgagcgcacc
 
-
tttaactggcaattaatgcaacaaattagtgggcaatgtcatttatatgaccccgaacat
 
-
ggctaccgcaccttcggctccctgaaaaaagtttaa</font></p></div>
 
-
  </ul>
 
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</div>
 
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<div class="btn-group">
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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    Δ15 Synechocystis
 
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    <span class="caret"></span>
 
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  </a>
 
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  <ul class="dropdown-menu">
 
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    <div class="span5"><p><font size="1">gtgcgtctagaaatttcatcgcctcaaacaaagcttccttaccccaaaactgaagaatta
 
-
ccatttaccctccaagagctcagaaacgctattccagcggattgttttgagccatcggta
 
-
gtccggtccttgggctacttttttttggatgttggtttaattgccgggttttatgctcta
 
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gcggcctaccttgattcctggttcttctatccgattttttggttaattcagggaacccta
 
-
ttctggtccctgtttgtggtgggccatgattgtggccatggctccttttccaaatccaaa
 
-
acccttaataattggattggtcatctcagccacacgccaattttggtgccttaccatggc
 
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tggcgtattagtcatcgtactcaccatgccaacacgggcaatatcgacaccgacgaaagt
 
-
tggtatccagtgtcggagcaaaaatataaccaaatggcctggtatgaaaaacttctacgt
 
-
ttttacttgcctctgatcgcctaccccatttatctatttcggcgatcgccaaaccggcaa
 
-
ggctcccatttcatgcccggcagtcccctattccgtcccggagaaaaagcagctgttctc
 
-
accagcacctttgcccttgcagcctttgtcggcttccttggctttttaacttggcaattt
 
-
ggctggctatttttgctgaaattttatgttgccccctacctcgtgtttgtggtgtggtta
 
-
gatttggtcacatttttacatcacactgaagacaatatcccttggtatcgtggtgatgac
 
-
tggtattttctcaaaggtgccctctccaccattgatcgggattacggcttcattaacccc
 
-
attcaccatgacattggcacccacgtcgcccaccatattttctcgaatatgccccactac
 
-
aagttacgccgggcgactgaagccatcaagcccattttaggggaatattatcgatattct
 
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gacgagccaatttggcaagctttttttaagtcctactgggcttgccattttgttcctaat
 
-
caaggttcaggggtctattaccaatccccatccaatggtggatatcaaaagaaaccttaa</font></p></div>
 
-
  </ul>
 
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</div>
 
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<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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    Δ6 Synechocystis
 
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    <span class="caret"></span>
 
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  </a>
 
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  <ul class="dropdown-menu">
 
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    <div class="span5"><p><font size="1">atgctaacagcggaaagaattaaatttacccagaaacgggggtttcgtcgggtactaaac
 
-
caacgggtggatgcctactttgccgagcatggcctgacccaaagggataatccctccatg
 
-
tatctgaaaaccctgattattgtgctctggttgttttccgcttgggcctttgtgcttttt
 
-
gctccagttatttttccggtgcgcctactgggttgtatggttttggcgatcgccttggcg
 
-
gccttttccttcaatgtcggccacgatgccaaccacaatgcctattcctccaatccccac
 
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atcaaccgggttctgggcatgacctacgattttgtcgggttatctagttttctttggcgc
 
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tatcgccacaactatttgcaccacacctacaccaatattcttggccatgacgtggaaatc
 
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catggagatggcgcagtacgtatgagtcctgaacaagaacatgttggtatttatcgtttc
 
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cagcaattttatatttggggtttatatcttttcattcccttttattggtttctctacgat
 
-
gtctacctagtgcttaataaaggcaaatatcacgaccataaaattcctcctttccagccc
 
-
ctagaattagctagtttgctagggattaagctattatggctcggctacgttttcggctta
 
-
cctctggctctgggcttttccattcctgaagtattaattggtgcttcggtaacctatatg
 
-
acctatggcatcgtggtttgcaccatctttatgctggcccatgtgttggaatcaactgaa
 
-
tttctcacccccgatggtgaatccggtgccattgatgacgagtgggctatttgccaaatt
 
-
cgtaccacggccaattttgccaccaataatcccttttggaactggttttgtggcggttta
 
-
aatcaccaagttacccaccatcttttccccaatatttgtcatattcactatccccaattg
 
-
gaaaatattattaaggatgtttgccaagagtttggtgtggaatataaagtttatcccacc
 
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ttcaaagcggcgatcgcctctaactatcgctggctagaggccatgggcaaagcatcgtga</font></p></div>
 
-
  </ul>
 
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</div>
 
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<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    ELO 6 L. major
 
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    <span class="caret"></span>
 
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  </a>
 
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  <ul class="dropdown-menu">
 
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    <div class="span5"><p><font size="1">atgaaggtcatcgtcgcttctggcccggacggtgcccgcaagcacgaggtggagctggca
 
-
gccaacgccacgctcgcagatctgaagaaggcctaccaacggggtgtggacgtgcaccgc
 
-
aagtcgttcaaggttcccagcgcggagtcgccgctgccaggtgcggatagtggcaagctg
 
-
cgcccgaacctcattactctgtcagataaggtgcccctgtcgcagcagggggtgaaggat
 
-
ggctcggtgatcacttacaaggacctcggcccgcagatcggctaccgcacggtgttctac
 
-
gtcgagtatgccggccccatcgccttcatgctgctgtacgccatgcgcccttcgctcatc
 
-
tacggctctgccccgatgccggcttacggctacacgcagaagctatacattggcctcttc
 
-
ctcgcccacttcttaaagcgcgagctcgagtccatgtttgtgcacaagttctcgcaccca
 
-
acgatgccgatgcgcaacatcttcaagaactgcatctactactggtccttcgccgccttc
 
-
atcggctacgtgctgtgcagtccttcattcacgccgaccagcaccatgcagtcaaacttc
 
-
ggcgccgtggtcatggtcatcaacgagctgctgaacttcgcggtgcactaccagcttagc
 
-
acgatgcgcaagtccgatggtgacaccacccgcaacgtgccgcaaggccctctgttcgcc
 
-
ttcgtctcgtgcccgaactacttctttgagattatgtcgtgggtgtccttttccatcggc
 
-
acaaatatgttatcctcctggttcttcacactcgccggtttcgtgcagatggcggactgg
 
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gcgaagaagaagcaccggggctacgtcaaggcggacccggccaataaaaagaaggccgcc
 
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attctgcccttcatcatgtag</font></p></div>
 
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  </ul>
 
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</div>
 
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<BR>&nbsp;</BR><BR>&nbsp;</BR><BR>&nbsp;</BR>
 
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<a name="Primers"><span class="label">Primers</span></a>
 
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<BR>&nbsp;</BR>
 
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<p>All primers are notated 5' to 3'.
 
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<BR>&nbsp;</BR>
 
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<div class="btn-toolbar">
 
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<div class="btn-group">
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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  Δ12 T. Cruzi forward
 
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    <span class="caret"></span>
 
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  </a>
 
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  <ul class="dropdown-menu">
 
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    <div class="span3">ATATCATATGTGGCGGGTTGACAATTTA
 
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  </ul>
 
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</div>
 
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<div class="btn-group">
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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    Δ12 T. Cruzi reverse
 
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    <span class="caret"></span>
 
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  </a>
 
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  <ul class="dropdown-menu">
 
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  <p><div class="span3">ATATCTCGAGTTACCCATGAAAGACAC</div></p> 
 
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  </ul>
 
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</div>
 
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<div class="btn-group">
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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    Δ12 Synechocystis forward
 
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    <span class="caret"></span>
 
-
  </a>
 
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  <ul class="dropdown-menu">
 
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      <div class="span3">ATATCATATGACTGCCACGATTCCC</div>
 
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  </ul>
 
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</div>
 
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<div class="btn-group">
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ12 Synechocystis reverse
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
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      <div class="span3">ATATCTCGAGTTAAACTTTTTTCAGGGAGC</div>
 
-
  </ul>
 
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</div>
 
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<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ15 Synechocystis forward
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
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      <div class="span3">ATATCATATCGCTCTAGAAATTTCATC</div>
 
-
  </ul>
 
-
</div>
 
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</div>
 
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<p>
 
-
<div class="btn-toolbar">
 
-
<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ15 Synechocystis reverse
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
-
      <div class="span3">ATATCACGACTTAAGGTTTCTTTTGATATC</div>
 
-
  </ul>
 
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</div>
 
-
 
-
<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ6 Synechocystis forward
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
-
<div class="span3">ATATCATATGCAAACAGCGGAAAGAATT</div>
 
-
  </ul>
 
-
</div>
 
-
 
-
<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ6 Synechocystis reverse
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
-
  <div class="span4">ATATCTCGAGTAACGATGCTTTGACCATGGCCTCTAG</div>
 
-
  </ul>
 
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</div></p>
 
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</div>
 
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<BR>&nbsp;<?BR>
 
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<p>When using the pET-duet vector, we needed additionally primers for the alternative restriction sites and His-tags.</p>
 
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<div class="btn-toolbar">
 
-
<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    ELO6 L. major forward
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
-
  <div class="span6">TATCATATGGGAAGCAGCCATCATCATCATCATCACAGCATGGTCTCTCTGGAGCAGGCC</div>
 
-
  </ul>
 
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</div>
 
-
 
-
<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ15 T. Cruzi forward
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
-
  <div class="span6">TATCATATGGGCAGCCATCATCATCATCATCACAGCATGCGTCTAGAAATTTCATCG</div>
 
-
  </ul>
 
-
</div>
 
-
 
-
<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ6 T. Cruzi forward
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
-
  <div class="span4">GGATCCGAATTCATCTAACAGCGGAAAGAATT</div>
 
-
  </ul>
 
-
</div>
 
-
 
-
<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ6 T. Cruzi reverse
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
-
  <div class="span4">GACAAGCTTTCACGATGCTTTGCCCATGGCCTCTAC</div>
 
-
  </ul>
 
-
</div>
 
-
 
-
<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ12 T. Cruzi forward
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
-
  <div class="span4"> GGATCCGAATTCATGACTGCCACGATTCCC</div>
 
-
  </ul>
 
-
</div>
 
-
 
-
<div class="btn-group">
 
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
-
    Δ12 T. Cruzi reverse
 
-
    <span class="caret"></span>
 
-
  </a>
 
-
  <ul class="dropdown-menu">
 
-
<div class="span3">GACAAGCTTTTAAACTTTTTTCAGGGAGC</div>
 
-
  </ul>
 
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</div>
 
-
</div>
 
-
<BR>&nbsp;</BR>
 
-
     
 
-
 
-
<a name="Sequencing"><span class="label">Sequencing results</span></a>
 
-
<BR>&nbsp;</BR>
 
-
 
-
 
-
 
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-
 
-
<dl class="dl-horizontal"> 
 
-
        <dt>date and gene</dt> 
 
-
        <dd>result</dd> 
 
-
        <dt>date and gene</dt> 
 
-
        <dd>result</dd>
 
-
        <dt>date and gene</dt> 
 
-
        <dd>result</dd>
 
-
        <dt>23/7/12 Δ12 Syn.</dt> 
 
-
        <dd><p>15b forward gave 0 nucleotides</p>
 
-
            <p>15b reverse</p>
 
-
            <p>20b forward gave a 41 nucleotide result.</p>
 
-
 
-
            <div class="btn-group">
 
-
            <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">Sequence<span class="caret"></span>
 
-
            </a>
 
-
              <ul class="dropdown-menu">
 
-
                  <div class="span5"><font size="1">gCGnCctGntGCCgCGCGGCnnnnnnnnnnnnnnnnnngac</font></div>
 
-
              </ul>
 
-
            </div>
 
-
           
 
-
            <p>A BLAST search showed this to be compatible with <i>Crocosphaera watsonii</i> (genome shotgun sequence, 21%), a diazotrophic cyanobacteria. Add in relations to synechocystis.</p>
 
-
 
-
            <p>20b reverse</p>
 
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<a name="Lipid"><span class="label">Lipid analysis</span></a>
 
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      <p>who knows what those Metal Mickies are up to. Eating nachos in the Whey Pat most likely.</p>
 
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Latest revision as of 11:51, 23 September 2012