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Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jun
From 2014.igem.org
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| - | <li>Transformation of electrocompotetent <i> E.coli </i> KRX cells:</li> | + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Transformationviaelectroporation" target="_blank">Transformation of electrocompotetent <i> E.coli </i> KRX cells:</a></li> |
<ul> | <ul> | ||
<li>K538742: <i>pSB1C3-kivD</i> (parts distribution from 2013, plate 1, 24L)</li> | <li>K538742: <i>pSB1C3-kivD</i> (parts distribution from 2013, plate 1, 24L)</li> | ||
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<li>incubation over night at 37°C and 250 rpm</li> | <li>incubation over night at 37°C and 250 rpm</li> | ||
</ul> | </ul> | ||
| - | <li>Plasmid isolation by | + | <li>Plasmid isolation by ThermoScientific MiniPrep</li> |
<li>Sanger sequencing using <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#VF-Primer" target="_blank">VF</a> and <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#VR-Primer" target="_blank">VR</a> as sequencing primers</li> | <li>Sanger sequencing using <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#VF-Primer" target="_blank">VF</a> and <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#VR-Primer" target="_blank">VR</a> as sequencing primers</li> | ||
<ul> | <ul> | ||
Revision as of 05:01, 27 August 2014
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June |
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- Transformation of electrocompotetent E.coli KRX cells:
- K538742: pSB1C3-kivD (parts distribution from 2013, plate 1, 24L)
- K539621: pSB1C3-ilvC (parts distribution from 2013, plate 1, 24F)
- K539626: pSB1C3-ilvD (parts distribution from 2013, plate 1, 22J)
- K539627: pSB1C3-alsS (parts distribution from 2013, plate 1, 24H)
- Resulting colonies were selected to start liquid cultures:
- 5 ml of LB in 20 ml reaction tube
- incubation over night at 37°C and 250 rpm
- Plasmid isolation by ThermoScientific MiniPrep
- Sanger sequencing using VF and VR as sequencing primers
- Identities of all parts were confirmed, but not whole sequence of the parts was covered
- Glycerin stocks were created for all four strains:
- E. coli KRX pSB1C3-alsS
- E. coli KRX pSB1C3-ilvC
- E. coli KRX pSB1C3-ilvD
- E. coli KRX pSB1C3-kivD
