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Team:Hannover/Results/Plant Vector
From 2014.igem.org
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<h1>Results / Plant vector</h1><p class="text">We want to provide iGEM a new Plant vector with a BioBrick <a href="http://parts.igem.org/Help:Standards/Assembly/RFC21">RFC[21]MCS</a> and 2x35S promotor.</p> | <h1>Results / Plant vector</h1><p class="text">We want to provide iGEM a new Plant vector with a BioBrick <a href="http://parts.igem.org/Help:Standards/Assembly/RFC21">RFC[21]MCS</a> and 2x35S promotor.</p> | ||
| - | <h2>Labwork</h2><p class="text"><ul><li>exchange the original promoter of the binary vector pORE_E3 <a href="http://www.ncbi.nlm.nih.gov/nuccore/AY562536">(AY562536.1)</a> into a 2x35s promoter using <a href="">this protocol</a></li><li><a href="">removal of restriction sites</a> in our pORE_E3_2x35S </li><li>cloning the RFC[21]MCS into this vector</li></ul></p> | + | <h2>Labwork</h2><p class="text"><ul><li>exchange the original promoter enTCUP2 of the binary vector pORE_E3 <a href="http://www.ncbi.nlm.nih.gov/nuccore/AY562536">(AY562536.1)</a> into a 2x35s promoter using <a href="">this protocol</a></li><li><a href="">removal of restriction sites</a> in our pORE_E3_2x35S </li><li>cloning the RFC[21]MCS into this vector</li></ul></p> |
<h2>Results</h2><p class="text">The exchange of the original promoter of the pORE_E3 vector was successful. We gained a “new” vector with a 2x35s promoter (pORE_E3_2x35s). Furthermore we were able to remove the restriction sites XhoI and BglII from this vector. Unfortunately our last attempt, changing the MCS to the MCS of RFC[21] via the insertion of two hybridized primers with overhangs, failed several times. Even a modified protocol couldn’t bring us the expected success. At the end, we ran out of time.<br><br>Of cause, we could have synthesized the insert, cut it with restriction enzymes and then clone it into the vector, but that’s what everyone could have done…</p> | <h2>Results</h2><p class="text">The exchange of the original promoter of the pORE_E3 vector was successful. We gained a “new” vector with a 2x35s promoter (pORE_E3_2x35s). Furthermore we were able to remove the restriction sites XhoI and BglII from this vector. Unfortunately our last attempt, changing the MCS to the MCS of RFC[21] via the insertion of two hybridized primers with overhangs, failed several times. Even a modified protocol couldn’t bring us the expected success. At the end, we ran out of time.<br><br>Of cause, we could have synthesized the insert, cut it with restriction enzymes and then clone it into the vector, but that’s what everyone could have done…</p> | ||
Revision as of 10:46, 15 October 2014
Results / Plant vector
We want to provide iGEM a new Plant vector with a BioBrick RFC[21]MCS and 2x35S promotor.
Labwork
- exchange the original promoter enTCUP2 of the binary vector pORE_E3 (AY562536.1) into a 2x35s promoter using this protocol
- removal of restriction sites in our pORE_E3_2x35S
- cloning the RFC[21]MCS into this vector
Results
The exchange of the original promoter of the pORE_E3 vector was successful. We gained a “new” vector with a 2x35s promoter (pORE_E3_2x35s). Furthermore we were able to remove the restriction sites XhoI and BglII from this vector. Unfortunately our last attempt, changing the MCS to the MCS of RFC[21] via the insertion of two hybridized primers with overhangs, failed several times. Even a modified protocol couldn’t bring us the expected success. At the end, we ran out of time.
Of cause, we could have synthesized the insert, cut it with restriction enzymes and then clone it into the vector, but that’s what everyone could have done…
