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Team:Warsaw/Calendar-Main/4 August 2008
From 2008.igem.org
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<h3> Cloning of truncated fragment of protein A</h3> | <h3> Cloning of truncated fragment of protein A</h3> | ||
<h4>Piotr</h4> | <h4>Piotr</h4> | ||
| - | <p>Inoculation of some pACYC177+OmpA_alpha + deltaA and pACYC177+OmpA_omega + deltaA colonies of tranformants.</p> | + | <p>Inoculation of some <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-deltaA-alpha>pACYC177+OmpA_alpha + deltaA</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BOmpA-deltaA-omega>pACYC177+OmpA_omega + deltaA</a> colonies of tranformants.</p> |
| - | <h3>Checking the expression of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha> | + | <h3>Checking the expression of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_ΔA_alpha</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-alpha>OmpA_ΔA_alpha</a></h3><h4>Piotr</h4> |
| - | <p>Inoculation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha> | + | <p>Inoculation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_ΔA_alpha</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-deltaA-alpha>OmpA_ΔA_alpha</a> with inductor (0,5 mmol/mL IPTG) and negative control without IPTG.</p> |
| - | <h3>Checking if | + | <h3>Checking if <a hres=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_ΔA_alpha</a> gives ampicillin resistance</h3><h4>Emilia</h4> |
| - | <p>Inoculation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha> | + | <p>Inoculation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_ΔA_alpha</a> into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75, 1 mmol/mL.</p> |
| - | <h3>Preparing pACYC177+OmpA_omega_deltaA construct</h3><h4>Michał K.</h4> | + | <h3>Preparing <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-deltaA>pACYC177+OmpA_omega_deltaA</a> construct</h3><h4>Michał K.</h4> |
| - | <ol><li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of pACYC177+OmpA_omega_deltaA_alpha (from 25 July) with BamHI and NotI (BamHI buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> with Klenow fragment (3 hr). </li> | + | <ol><li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>pACYC177+OmpA_omega_deltaA_alpha</a> (from 25 July) with BamHI and NotI (BamHI buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> with Klenow fragment (3 hr). </li> |
<li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 4300 bp. </li> | <li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 4300 bp. </li> | ||
<li>Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragment. </li> | <li>Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragment. </li> | ||
Revision as of 15:16, 25 October 2008
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Cloning of truncated fragment of protein APiotrInoculation of some pACYC177+OmpA_alpha + deltaA and pACYC177+OmpA_omega + deltaA colonies of tranformants. Checking the expression of OmpA_omega_ΔA_alpha and OmpA_ΔA_alphaPiotrInoculation of OmpA_omega_ΔA_alpha and OmpA_ΔA_alpha with inductor (0,5 mmol/mL IPTG) and negative control without IPTG. Checking if OmpA_omega_ΔA_alpha gives ampicillin resistanceEmiliaInoculation of OmpA_omega_ΔA_alpha into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75, 1 mmol/mL. Preparing pACYC177+OmpA_omega_deltaA constructMichał K.
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