From 2008.igem.org
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| - | <h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA</h3>
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| - | <h4>Paweł</h4>
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| - | <li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from cultures inocluated on previous day</li>
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| - | <li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmids with NdeI and NotI. Two positives obtained (proper band: ~150 bp visible)</li>
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| - | <li>One of positive plasmids transformed into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB+amp, overnight, for further isolation of pET15b+Z+omega vector</li>
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| | + | <h3>Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA</h3><h4>Paweł</h4> |
| | + | <ol><li>Results of ligation: 6 colonies grown.</li> |
| | + | <li>Each colony cultured overnight in LB + ampicillin.</li></ol> |
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Revision as of 20:20, 11 October 2008
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Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpAPaweł
- Results of ligation: 6 colonies grown.
- Each colony cultured overnight in LB + ampicillin.
- Separate transformant colonies (tranformation of ligation of pACYC177+OmpA_alpha and omega_A from previous day) inoculated to liquid LB with kanamycin.
Antoni:
- Preparation of chemocompetent bacteria E. coli TOP10.
Cloning omega-A fusion on pKS (second attempt)
Michał L., Ewa, Marcin:
Sequencing confirms that we have obtained proper construct with omega-A fusion
Cloning of protein A DNA to OmpA constructs
- Colony PCR on colonies from plates with transformations pACYC177+OmpA_A_omega. Primers used:
AL+SacI and AP+NotI
- Confirmed transformant colonies inoculated to liquid LB with kanamycin.
- Isolation of plasmids from cultures inocluated on previous day (pACYC177+OmpA_A_alpha).
- Control digest of isolated plasmids with BamHI and SacI.
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