Team:St Andrews/Procedure

From 2012.igem.org

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<p>For primer annealing in the PCR, the primer sequences were combined in the following way:</p>
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<li>GST Forward and Ni Reverse</li>
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<li>GST Forward + Ni<sub>2</sub> Reverse</li>
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<li>GST Forward + Pd Reverse</li>
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<li>GST Forward + Pt Reverse</li>
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Revision as of 14:40, 27 July 2012

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 Δ6 Synechocystis ELO 6 L. major
 

 
Primers
 

All primers are notated 5' to 3'.
 

Δ12 T. Cruzi forward
Δ12 T. Cruzi reverse
Δ12 Synechocystis forward
Δ12 Synechocystis reverse
Δ15 Synechocystis forward

Δ15 Synechocystis reverse
Δ6 Synechocystis forward
Δ6 Synechocystis reverse


 

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

ELO6 L. major forward
Δ15 T. Cruzi forward
Δ6 T. Cruzi forward
Δ6 T. Cruzi reverse
Δ12 T. Cruzi forward
Δ12 T. Cruzi reverse

 
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 gave 0 nucleotides

20b forward gave a 41 nucleotide result.

Sequence

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 gave 0 nucleotides


 

 
Lipid analysis
 

 

 

 

 

 

 

 

 

who knows what those Metal Mickies are up to. Eating nachos in the Whey Pat most likely. MEGA LOLS (all caps, with a Z).


 

Primers

Primers
 

All primers are notated 5' to 3'.
 

Ni forward
Ni reverse
Ni2 forward
Ni2 reverse
.

Pd forward
Pd reverse
Pt forward
Pt reverse

For primer annealing in the PCR, the primer sequences were combined in the following way:

  • GST Forward and Ni Reverse
  • GST Forward + Ni2 Reverse
  • GST Forward + Pd Reverse
  • GST Forward + Pt Reverse

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    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.