Team:UPO-Sevilla/Notebook/08 10

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       <h1>August, 10th</h1>
       <h1>August, 10th</h1>
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       <h2>Assembly Team</h2>
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       <h2>Production Team</h2>
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      <p>We repeated SDM reactions for <i>fecA*</i> (UPO28) and <i>gltD**</i> (UPO17) from the beginning. This time we used <i>Taq expand enzyme</i> instead of <i>Pfu</i>. In the results, we did not obtain any product of amplification in the first SDM’s PCR reaction. Next we made the second SDM’s PCR reaction using first former SDM’s PCR reaction products. Again the results were not satisfactory.</p>
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<p>We repeated SDM reactions for <i>fecA*</i> (UPO28) and <i>gltD**</i> (UPO17) from the beginning. This time we used <i>Taq expand enzyme</i> instead of <i>Pfu</i>. In the results, we did not obtain any product of amplification in the first PCR reaction of SDM. Next we made the second PCR reaction of SDM by using the former PCR reaction products of SDM. Again the results were not satisfactory.</p>
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      <h2>Assembly Team</h2>
       <p>Transformation of <i>E. coli</i> DH5α with ligations made the day before.</p>
       <p>Transformation of <i>E. coli</i> DH5α with ligations made the day before.</p>
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       <p>We analyzed BamI-PstI digestions in 1% agarose gel: colony 4 and 5 are positive.</p>
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       <p>BamI-PstI digestions were analyzed by 1% agarose electrophoresis gel: colony 4 and 5 were positive.</p>
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       <p>The 2+28 and 28+3 day before transformation plates have not white colonies. 7+2+28 colonies are strange. We will repeat these devices using low copy vectors. We set up inocula of positive candidate colonies.</p>
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       <p>The 2+28 and 28+3 day before transformation plates had not white colonies. Colonies from UPO7+2+28 plate were strange. We will repeat these devices using low copy vectors. We set up inocula of positive candidate colonies.</p>
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       <p>Preparative digestion of UPO 4, 5 and 11 (in Mr. Gene vector) and 1% agarose gel. GFX purification. Ligation with pSB1C3 and transformation.</p>
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       <p>We made preparative digestion of UPO 4, 5 and 11 (in Mr. Gene vector) and ran them in 1% agarose gel. The spots were purified using GFX. Ligations with pSB1C3 and transformations were made.</p>

Latest revision as of 00:26, 27 October 2010

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August, 10th

Production Team

We repeated SDM reactions for fecA* (UPO28) and gltD** (UPO17) from the beginning. This time we used Taq expand enzyme instead of Pfu. In the results, we did not obtain any product of amplification in the first PCR reaction of SDM. Next we made the second PCR reaction of SDM by using the former PCR reaction products of SDM. Again the results were not satisfactory.

Assembly Team

Transformation of E. coli DH5α with ligations made the day before.

BamI-PstI digestions were analyzed by 1% agarose electrophoresis gel: colony 4 and 5 were positive.

The 2+28 and 28+3 day before transformation plates had not white colonies. Colonies from UPO7+2+28 plate were strange. We will repeat these devices using low copy vectors. We set up inocula of positive candidate colonies.

We made preparative digestion of UPO 4, 5 and 11 (in Mr. Gene vector) and ran them in 1% agarose gel. The spots were purified using GFX. Ligations with pSB1C3 and transformations were made.

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