Team:Warsaw/Calendar-Main/18 September 2008
From 2008.igem.org
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- | <h3>Preparation of | + | <h3>Preparation of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+Omp_</h3> |
<h4>Michał K.</h4> | <h4>Michał K.</h4> | ||
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- | <li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of | + | <li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+OmpA_alpha</a> plasmid with NdeI and SacI (BamHI buffer). <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">Dephosphorylation</a> (CIAP) of plasmid.</li> |
- | <li> Gel electrophoresis of digested | + | <li> Gel electrophoresis of digested plasmid |
- | + | and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands (OmpA_link - 500 bp)(<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/18_September_2008#fig4">Fig. 4.</a>).</li> | |
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+ | <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/18_September_2008#fig5">Fig. 5</a> | ||
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/c/c3/Grad2_17_09.jpg" width=300/></a><var><b>Fig. 1. Gradient PCR (temperatures:55-75 °C)</b><br> | <a name="fig1"><img src="https://static.igem.org/mediawiki/2008/c/c3/Grad2_17_09.jpg" width=300/></a><var><b>Fig. 1. Gradient PCR (temperatures:55-75 °C)</b><br> |
Revision as of 00:06, 27 October 2008
'Hunter and prey' system tests: Competition testsWeronikaInoculation of pACYC177+OmpA_A_alpha, pACYC177+OmpA_Z_omega, pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_alpha + induction using 0.25mM iPTG MutD5 testingPiotrInoculation of mutD5 into 100 ml of LB and rendering them chemocompetent. Electroporation with pACYC177+OmpA_Z_alpha, pACYC177+OmpA_Z_omega, pACYC177+OmpA_A_omega and pACYC177+OmpA_omega_ΔA. Optimisation of primers for preparation of partsMichał K.
Preparation of pSB1A3 carrying ΔA (BBa_K103003)Michał K.
Preparation of pSB2K3 + linker_alpha (BBa_K103009)Michał K.
Preparation of pSB2K3 + linker_omega (BBa_K103013)Michał K.
Preparation of pACYC177 + OmpA_omega_Michał K.
Preparation of pSB1A3+Omp_Michał K.
1. Marker 2. 55 °C linker_alpha 3. 60 °C linker_alpha 4. 65 °C linker_alpha 5. 70 °C linker_alpha Fig. 2. PCR to obtain inserts 1. Marker 2. deltaA 3 omega 4. ompA_omega Fig. 3. PCR to obtain linker_alpha 1. Marker 2. linker_alpha Fig. 4. Control SacI/NdeI digest of pET15b_OmpA_omega 1. Marker 2. digested pSB1A3 Fig. 5. Control EcoRI/BcuI digest of pSB1A3 1. Marker 2. digested pSB1A3
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