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Team:Valencia/Parts
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| - | + | ==Construction of Valencia Team Biobricks== | |
| + | |||
| + | ===Preparing inserts=== | ||
| + | |||
| + | 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> | ||
| + | Forward: 5' '''gAATTC'''gCggCCgCTTCTAgATggTgAgTTCgACAACTTC 3';<br> | ||
| + | Reverse: 5' TACTAgTAgCggCCg'''CTgCAg'''CTATgTggTgCAgTCCACTg 3' <br> | ||
| + | |||
| + | |||
| + | PCR was conducted as follows:<br> | ||
| + | |||
| + | <ol>A first denaturation cycle | ||
| + | <ol>94º 3'</ol> | ||
| + | Followed by 30 amplification cycles: | ||
| + | <ol>94º 30'' | ||
| + | 60º 1' 30'' | ||
| + | 72º 1'</ol> | ||
| + | And a final extension step: | ||
| + | <ol>72º 10'</ol> | ||
| + | </ol> | ||
</div> | </div> | ||
</div> | </div> | ||
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'
