Team:Warsaw/Calendar-Main/27 May 2008

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<html><h3>Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPMT5-AID+T7</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPMT5+AID-T7</a></h3>
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<h4>Michał K.</h4>
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<p> Further inoculation of separate transformant colonies for transcriptional and translational fusions on liquid LB + tetracycline (as there were no positive colonies in previous assays).
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1. Isolation of hypothetical pMPM-T5 with transcription fusion
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2. Isolation of hypothetical pMPM-T5 with translation fusion
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3. Control digest of pMPM-T5 with transcription fusion with EcoRI and XhoI
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4. Control digest of pMPM-T5 with translation fusion with BamHI
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5. Control digest of pMPM-T5 with translation fusion with EcoRI and XbaI
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6. Choice of proper clones and preparation for sequencing
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<h3>Here we found the clone with translational fusion on pMPM. These are different isolations in each lane. We have been working with the AT6 ever since. <br>
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Digestion was conducted with BamHI</h3>
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<center>https://static.igem.org/mediawiki/2008/6/62/Trawienie_06_05_2008.jpg</center>
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Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Michał K.

Further inoculation of separate transformant colonies for transcriptional and translational fusions on liquid LB + tetracycline (as there were no positive colonies in previous assays).




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