Team:St Andrews/Procedure

From 2012.igem.org

(Difference between revisions)
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<BR>&nbsp;</BR>
<BR>&nbsp;</BR>
 +
<a name="Sequences"><span class="label">Sequences</span></a>
<a name="Sequences"><span class="label">Sequences</span></a>
 +
 +
<p>
<div class="btn-toolbar">
<div class="btn-toolbar">
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<div class="btn-group">
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<a data-toggle="modal" href="#modal1" class="btn btn-primary btn-small">Δ12 T. Cruzi</a>
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
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<div class="modal hide" id="modal1">
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    Δ12 T. Cruzi
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  <div class="modal-header">
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     <span class="caret"></span>
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     <button type="button" class="close" data-dismiss="modal">×</button>
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   </a>
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    <h3>Δ12 T. Cruzi</h3>
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   <ul class="dropdown-menu">
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   </div>
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     <div class="span5"><p><font size="1">
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   <div class="modal-body">
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atgtcgcgggttgacaatttaactgtcgcgccgggtccaccggatgtcatgaaggcggtgcttaagtctgggcgcaacaaggagcacaat
+
     <p>atgtcgcgggttgacaatttaactgtcgcgccgggtccaccggatgtcatgaaggcggtgcttaagtctgggcgcaacaaggagcacaat
gtcattgtgccttcctccgcactgaccattggagaaattcaggagaaaatcccggtaaagtactttgaacgcaacactacccggagcgtc
gtcattgtgccttcctccgcactgaccattggagaaattcaggagaaaatcccggtaaagtactttgaacgcaacactacccggagcgtc
atgtttcttttacgcgacctcgcacaggtggcgttgacgtacgccattatgtacgccgtcgccttgccacttgccacttccatggaagtttctg
atgtttcttttacgcgacctcgcacaggtggcgttgacgtacgccattatgtacgccgtcgccttgccacttgccacttccatggaagtttctg
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acatggctccatcacatcaatgattcccacgtggtacatcacctctttagcaaaatgccacactacaacgcaatccaggtgacaagaaag
acatggctccatcacatcaatgattcccacgtggtacatcacctctttagcaaaatgccacactacaacgcaatccaggtgacaagaaag
tacattcgtgagatactgggtgccacatacattacggatgagaggtcactgtggaagatgctctgggaacagcgtagagagtgccgctatg
tacattcgtgagatactgggtgccacatacattacggatgagaggtcactgtggaagatgctctgggaacagcgtagagagtgccgctatg
-
ttgttcccgcagagggcgtctgtgtctttcatgggtaa</font></p></div>
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ttgttcccgcagagggcgtctgtgtctttcatgggtaa</p>
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   </ul>
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  </div>
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   <div class="modal-footer">
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    <a href="#" class="btn" data-dismiss="modal">Close</a>
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    </div>
</div>
</div>
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<div class="btn-group">
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<a data-toggle="modal" href="#modal1" class="btn btn-primary btn-small">Δ12 Synechocystis</a>
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
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<div class="modal hide" id="modal1">
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    Δ12 Synechocystis
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  <div class="modal-header">
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     <span class="caret"></span>
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     <button type="button" class="close" data-dismiss="modal">×</button>
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   </a>
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    <h3>Δ12 Synechocystis</h3>
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   <ul class="dropdown-menu">
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   </div>
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     <div class="span5"><p><font size="1">atgactgccacgattcccccgttgacaccaacggtaacgcccagcaatcccgatcgcccg
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   <div class="modal-body">
 +
     <p>atgactgccacgattcccccgttgacaccaacggtaacgcccagcaatcccgatcgcccg
attgcggatctcaaactacaagacatcattaaaaccctgcccaaggaatgcttcgagaaa
attgcggatctcaaactacaagacatcattaaaaccctgcccaaggaatgcttcgagaaa
aaagcgagcaaagcctgggcttctgttttgattaccctaggggcgatcgccgtgggctat
aaagcgagcaaagcctgggcttctgttttgattaccctaggggcgatcgccgtgggctat
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ctacgactagcccacggaagtttaaaagaaaactggggaccttttctttacgagcgcacc
ctacgactagcccacggaagtttaaaagaaaactggggaccttttctttacgagcgcacc
tttaactggcaattaatgcaacaaattagtgggcaatgtcatttatatgaccccgaacat
tttaactggcaattaatgcaacaaattagtgggcaatgtcatttatatgaccccgaacat
-
ggctaccgcaccttcggctccctgaaaaaagtttaa</font></p></div>
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ggctaccgcaccttcggctccctgaaaaaagtttaa
-
   </ul>
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</p>
 +
  </div>
 +
   <div class="modal-footer">
 +
    <a href="#" class="btn" data-dismiss="modal">Close</a>
 +
    </div>
</div>
</div>
-
<div class="btn-group">
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<a data-toggle="modal" href="#modal1" class="btn btn-primary btn-small">Δ15 Synechocystis</a>
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
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<div class="modal hide" id="modal1">
-
    Δ15 Synechocystis
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  <div class="modal-header">
-
     <span class="caret"></span>
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     <button type="button" class="close" data-dismiss="modal">×</button>
-
   </a>
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    <h3>Δ15 Synechocystis</h3>
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   <ul class="dropdown-menu">
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   </div>
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     <div class="span5"><p><font size="1">gtgcgtctagaaatttcatcgcctcaaacaaagcttccttaccccaaaactgaagaatta
+
   <div class="modal-body">
 +
     <p>gtgcgtctagaaatttcatcgcctcaaacaaagcttccttaccccaaaactgaagaatta
ccatttaccctccaagagctcagaaacgctattccagcggattgttttgagccatcggta
ccatttaccctccaagagctcagaaacgctattccagcggattgttttgagccatcggta
gtccggtccttgggctacttttttttggatgttggtttaattgccgggttttatgctcta
gtccggtccttgggctacttttttttggatgttggtttaattgccgggttttatgctcta
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aagttacgccgggcgactgaagccatcaagcccattttaggggaatattatcgatattct
aagttacgccgggcgactgaagccatcaagcccattttaggggaatattatcgatattct
gacgagccaatttggcaagctttttttaagtcctactgggcttgccattttgttcctaat
gacgagccaatttggcaagctttttttaagtcctactgggcttgccattttgttcctaat
-
caaggttcaggggtctattaccaatccccatccaatggtggatatcaaaagaaaccttaa</font></p></div>
+
caaggttcaggggtctattaccaatccccatccaatggtggatatcaaaagaaaccttaa
-
   </ul>
+
</p>
 +
  </div>
 +
   <div class="modal-footer">
 +
    <a href="#" class="btn" data-dismiss="modal">Close</a>
 +
    </div>
</div>
</div>
-
 
+
<a data-toggle="modal" href="#modal1" class="btn btn-primary btn-small">Δ12 Synechocystis</a>
-
<div class="btn-group">
+
<div class="modal hide" id="modal1">
-
  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
+
  <div class="modal-header">
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    Δ6 Synechocystis
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     <button type="button" class="close" data-dismiss="modal">×</button>
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     <span class="caret"></span>
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    <h3>Δ6 Synechocystis</h3>
-
   </a>
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   </div>
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   <ul class="dropdown-menu">
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   <div class="modal-body">
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     <div class="span5"><p><font size="1">atgctaacagcggaaagaattaaatttacccagaaacgggggtttcgtcgggtactaaac
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     <p>atgctaacagcggaaagaattaaatttacccagaaacgggggtttcgtcgggtactaaac
caacgggtggatgcctactttgccgagcatggcctgacccaaagggataatccctccatg
caacgggtggatgcctactttgccgagcatggcctgacccaaagggataatccctccatg
tatctgaaaaccctgattattgtgctctggttgttttccgcttgggcctttgtgcttttt
tatctgaaaaccctgattattgtgctctggttgttttccgcttgggcctttgtgcttttt
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aatcaccaagttacccaccatcttttccccaatatttgtcatattcactatccccaattg
aatcaccaagttacccaccatcttttccccaatatttgtcatattcactatccccaattg
gaaaatattattaaggatgtttgccaagagtttggtgtggaatataaagtttatcccacc
gaaaatattattaaggatgtttgccaagagtttggtgtggaatataaagtttatcccacc
-
ttcaaagcggcgatcgcctctaactatcgctggctagaggccatgggcaaagcatcgtga</font></p></div>
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ttcaaagcggcgatcgcctctaactatcgctggctagaggccatgggcaaagcatcgtga
-
   </ul>
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</p>
 +
  </div>
 +
   <div class="modal-footer">
 +
    <a href="#" class="btn" data-dismiss="modal">Close</a>
 +
    </div>
</div>
</div>
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<div class="btn-group">
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<a data-toggle="modal" href="#modal1" class="btn btn-primary btn-small">ELO 6 L. major</a>
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   <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
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<div class="modal hide" id="modal1">
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     ELO 6 L. major
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   <div class="modal-header">
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    <span class="caret"></span>
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    <button type="button" class="close" data-dismiss="modal">×</button>
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   </a>
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     <h3>ELO 6 L. major</h3>
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   <ul class="dropdown-menu">
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   </div>
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    <div class="span5"><p><font size="1">atgaaggtcatcgtcgcttctggcccggacggtgcccgcaagcacgaggtggagctggca
+
   <div class="modal-body">
 +
    <p>atgaaggtcatcgtcgcttctggcccggacggtgcccgcaagcacgaggtggagctggca
gccaacgccacgctcgcagatctgaagaaggcctaccaacggggtgtggacgtgcaccgc
gccaacgccacgctcgcagatctgaagaaggcctaccaacggggtgtggacgtgcaccgc
aagtcgttcaaggttcccagcgcggagtcgccgctgccaggtgcggatagtggcaagctg
aagtcgttcaaggttcccagcgcggagtcgccgctgccaggtgcggatagtggcaagctg
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acaaatatgttatcctcctggttcttcacactcgccggtttcgtgcagatggcggactgg
acaaatatgttatcctcctggttcttcacactcgccggtttcgtgcagatggcggactgg
gcgaagaagaagcaccggggctacgtcaaggcggacccggccaataaaaagaaggccgcc
gcgaagaagaagcaccggggctacgtcaaggcggacccggccaataaaaagaaggccgcc
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attctgcccttcatcatgtag</font></p></div>
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attctgcccttcatcatgtag</p>
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   </ul>
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  </div>
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   <div class="modal-footer">
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    <a href="#" class="btn" data-dismiss="modal">Close</a>
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    </div>
</div>
</div>
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<BR>&nbsp;</BR><BR>&nbsp;</BR><BR>&nbsp;</BR>
<BR>&nbsp;</BR><BR>&nbsp;</BR><BR>&nbsp;</BR>
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             <p>20b reverse</p>
             <p>20b reverse</p>
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            <a class="btn" data-toggle="modal" href="#myModal" >Launch Modal</a>
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         </dd>  
         </dd>  
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<BR>&nbsp;</BR><BR>&nbsp;</BR><BR>&nbsp;</BR><BR>&nbsp;</BR><BR>&nbsp;</BR>
<BR>&nbsp;</BR><BR>&nbsp;</BR><BR>&nbsp;</BR><BR>&nbsp;</BR><BR>&nbsp;</BR>
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</div>
</div>
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<div class="modal hide" id="myModal">
 
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    <button type="button" class="close" data-dismiss="modal">×</button>
 
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    <h3>Modal header</h3>
 
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  </div>
 
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  <div class="modal-body">
 
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    <p>One fine body…</p>
 
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  </div>
 
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  <div class="modal-footer">
 
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    <a href="#" class="btn" data-dismiss="modal">Close</a>
 
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    <a href="#" class="btn btn-primary">Save changes</a>
 
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  </div>
 
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</div>
 

Revision as of 10:25, 27 July 2012

St Andrews iGEM '12

Lab Book

Procedure

date
what we did
date
what we did
date
what we did

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.

Please also refer to our Protocols page.


 

 

 

 

 

 

 

 

 

 

 
Sequences   Primers   Sequencing results   Lipid analysis
 

Start from lipid extraction

primers, sequence results


 
Sequences

Δ12 T. Cruzi Δ12 Synechocystis Δ15 Synechocystis Δ12 Synechocystis ELO 6 L. major
 

 

 
Primers
 

All primers are notated 5' to 3'.
 

Δ12 T. Cruzi forward
    ATATCATATGTGGCGGGTTGACAATTTA
Δ12 T. Cruzi reverse

    ATATCTCGAGTTACCCATGAAAGACAC

Δ12 Synechocystis forward
    ATATCATATGACTGCCACGATTCCC
Δ12 Synechocystis reverse
    ATATCTCGAGTTAAACTTTTTTCAGGGAGC
Δ15 Synechocystis forward
    ATATCATATCGCTCTAGAAATTTCATC

Δ15 Synechocystis reverse
    ATATCACGACTTAAGGTTTCTTTTGATATC
Δ6 Synechocystis forward
    ATATCATATGCAAACAGCGGAAAGAATT
Δ6 Synechocystis reverse
    ATATCTCGAGTAACGATGCTTTGACCATGGCCTCTAG


 

When using the pET-duet vector, we needed additionally primers for the alternative restriction sites and His-tags.

ELO6 L. major forward
    TATCATATGGGAAGCAGCCATCATCATCATCATCACAGCATGGTCTCTCTGGAGCAGGCC
Δ15 T. Cruzi forward
    TATCATATGGGCAGCCATCATCATCATCATCACAGCATGCGTCTAGAAATTTCATCG
Δ6 T. Cruzi forward
    GGATCCGAATTCATCTAACAGCGGAAAGAATT
Δ6 T. Cruzi reverse
    GACAAGCTTTCACGATGCTTTGCCCATGGCCTCTAC
Δ12 T. Cruzi forward
    GGATCCGAATTCATGACTGCCACGATTCCC
Δ12 T. Cruzi reverse
    GACAAGCTTTTAAACTTTTTTCAGGGAGC

 
Sequencing results
 
date and gene
result
date and gene
result
date and gene
result
23/7/12 Δ12 Syn.

15b forward gave 0 nucleotides

15b reverse

20b forward gave a 41 nucleotide result.

Sequence
    gCGnCctGntGCCgCGCGGCnnnnnnnnnnnnnnnnnngac

A BLAST search showed this to be compatible with Crocosphaera watsonii (genome shotgun sequence, 21%), a diazotrophic cyanobacteria. Add in relations to synechocystis.

20b reverse


 

 
Lipid analysis
 

 

 

 

 

 

 

 

 

who knows what those Metal Mickies are up to. Eating nachos in the Whey Pat most likely.

Back to top

University of St Andrews, 2012.

Contact us: igem2012@st-andrews.ac.uk, Twitter, Facebook

This iGEM team has been funded by the MSD Scottish Life Sciences Fund. The opinions expressed by this iGEM team are those of the team members and do not necessarily represent those of Merck Sharp & Dohme Limited, nor its Affiliates.

Retrieved from "http://2012.igem.org/Team:St_Andrews/Procedure"