Team:Valencia/Parts

From 2008.igem.org

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[[Image:Valencia_UCP.png|600px|center| UCP function in mitochondria]]
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==Construction of Valencia Team Biobricks==
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===Preparing inserts===
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 +
Total DNA was extracted from our yeast strains.<br>
 +
UCP-1 was amplified by PCR using oligonucleotides matching the sequence and bearing the appropriate Biobrick prefix and suffix.<br>
 +
Primers' sequences (EcoRI and PstI sites in bold):<br>
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Forward: 5' '''gAATTC'''gCggCCgCTTCTAgATggTgAgTTCgACAACTTC 3';<br>   
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Reverse: 5' TACTAgTAgCggCCg'''CTgCAg'''CTATgTggTgCAgTCCACTg 3' <br>
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PCR was conducted as follows:<br>
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<ol>A first denaturation cycle
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<ol>94º 3'</ol>
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Followed by 30 amplification cycles:
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<ol>94º 30''
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60º 1' 30''
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72º 1'</ol>
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And a final extension step:
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<ol>72º 10'</ol>
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</ol>
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Revision as of 18:26, 29 October 2008



Construction of Valencia Team Biobricks

Preparing inserts

Total DNA was extracted from our yeast strains.
UCP-1 was amplified by PCR using oligonucleotides matching the sequence and bearing the appropriate Biobrick prefix and suffix.
Primers' sequences (EcoRI and PstI sites in bold):
Forward: 5' gAATTCgCggCCgCTTCTAgATggTgAgTTCgACAACTTC 3';
Reverse: 5' TACTAgTAgCggCCgCTgCAgCTATgTggTgCAgTCCACTg 3'


PCR was conducted as follows:

    A first denaturation cycle
      94º 3'

    Followed by 30 amplification cycles:

      94º 30 60º 1' 30 72º 1'

    And a final extension step:

      72º 10'