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Team:Warsaw/Calendar-Main/4 August 2008
From 2008.igem.org
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<h3>Preparing pACYC177+OmpA_omega_ΔA construct</h3><h4>Michał K.</h4> | <h3>Preparing pACYC177+OmpA_omega_ΔA construct</h3><h4>Michał K.</h4> | ||
<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_ΔA_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> | <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_ΔA_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 (Fig. 1 | + | <li>Gel elctrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/2_August_2008#fig1">Fig. 1</a>)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> | ||
</ol> | </ol> | ||
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| - | <img src="https://static.igem.org/mediawiki/2008/1/17/Gucio.jpg" width=240/> <var><b>Fig. 1. Gel-out of pACYC177_OmpA_omega_deltaA_alpha digested with NotI/BamHI after blunting ends.</b><br> | + | <a name="fig1"><img src="https://static.igem.org/mediawiki/2008/1/17/Gucio.jpg" width=240/></a> <var><b>Fig. 1. Gel-out of pACYC177_OmpA_omega_deltaA_alpha digested with NotI/BamHI after blunting ends.</b><br> |
1. Marker<br> | 1. Marker<br> | ||
2. pACYC177_OmpA_omega_deltaA_alpha <br></var> | 2. pACYC177_OmpA_omega_deltaA_alpha <br></var> | ||
Revision as of 17:34, 26 October 2008
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Cloning of truncated fragment of protein APiotrInoculation of some ACYC177+OmpA_alpha + ΔA and pACYC177+OmpA_omega + ΔA colonies of tranformants. Checking the expression of OmpA_omega_ΔA_alpha and OmpA_A_alphaPiotrInoculation of OmpA_omega_ΔA_alpha and OmpA_A_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_ΔA constructMichał K.
 Fig. 1. Gel-out of pACYC177_OmpA_omega_deltaA_alpha digested with NotI/BamHI after blunting ends.1. Marker 2. pACYC177_OmpA_omega_deltaA_alpha
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