Team:Warsaw/Calendar-Main/29 September 2008
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<h4>Piotr</h4> | <h4>Piotr</h4> | ||
<p>Sequencing results: there weren't any mutations in plasmids isolated from MutD<sub>5</sub> - maybe this strain is too weak as a mutator.</p> | <p>Sequencing results: there weren't any mutations in plasmids isolated from MutD<sub>5</sub> - maybe this strain is too weak as a mutator.</p> | ||
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+ | <h3>Preparation of pACYC177 + OmpA_omega_</h3> | ||
+ | <h4>Michał K.</h4> | ||
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+ | <ol> | ||
+ | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>pACYC177+OmpA_omega_ΔA_alpha</a> plasmid using | ||
+ | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmLNXNEB">OmLNXNEB</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinPBSNP">LinPBSNP</a> | ||
+ | primers (annealing temperature 58 °C; elongation length 90s) to obtain OmpA_omega fragment. </li> | ||
+ | <li> Gel electrophoresis of PCR products 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_omega_ - 900 bp).</li> | ||
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+ | </ol> | ||
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<li> Gel electrophoresis to estimate concentration of purified DNA from previous day. </li> | <li> Gel electrophoresis to estimate concentration of purified DNA from previous day. </li> | ||
<li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: <A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103004>Z (BBa_K103004)</a> and <A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103006>OmpA_linker (BBa_K103006)</a> (from 18 September). </li> | <li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: <A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103004>Z (BBa_K103004)</a> and <A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> + <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103006>OmpA_linker (BBa_K103006)</a> (from 18 September). </li> | ||
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>pACYC177+OmpA_omega_ΔA_alpha</a> plasmid using | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>pACYC177+OmpA_omega_ΔA_alpha</a> plasmid using | ||
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#PlacL_XNE">PlacL_XNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinP_BS">LinP_BS</a> | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#PlacL_XNE">PlacL_XNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinP_BS">LinP_BS</a> |
Revision as of 23:08, 26 October 2008
MutD5 testingPiotrSequencing results: there weren't any mutations in plasmids isolated from MutD5 - maybe this strain is too weak as a mutator. Preparation of pACYC177 + OmpA_omega_Michał K.
Preparation of BioBricksMichał K.
Preparation of vectors for BiobricksPiotrInoculation of bacteria received from iGEM HQs, carrying pSB2K3 and BBa_I739204 (pACYC177 converted into BioBrick vector) plasmids.
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