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Team:York UK/Notebook.html
From 2013.igem.org
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| - | <div id="wetlabcontent"><p><h2>Overview</h2> <p>My part of the team - Gintare, Evaldas and Jonas - were responsible for cloning of the mtr complex, gold sensing device and preparing peptides for expression.</p><h2>July 7th</h2> <p>Making of chemically competent DH5α cells. Protocol can be found in our protocols section. </p><h2>July 10th</h2> <p>Purification of genomic DNA from <i>Shewanella oneidensis</i> and <i>Salmonella sp.</i></p><h2>July 25th</h2> <p>After a long break we finally started working on the actual assembly of parts - the mtr complex. Today Jonas ran a trial PCR on mtrB and mtrC part of the complex. GoTaq polymerase (Promega) was used, this is the polymerase we generally use for trials and colony PCRs as fidelity is not the main priority.<img src="https://static.igem.org/mediawiki/2013/c/cc/Gel-mtrBC.jpg" width="340" height="150" alt="mtrBCtrial" align="left" | + | <div id="wetlabcontent"><p><h2>Overview</h2> <p>My part of the team - Gintare, Evaldas and Jonas - were responsible for cloning of the mtr complex, gold sensing device and preparing peptides for expression.</p><h2>July 7th</h2> <p>Making of chemically competent DH5α cells. Protocol can be found in our protocols section. </p><h2>July 10th</h2> <p>Purification of genomic DNA from <i>Shewanella oneidensis</i> and <i>Salmonella sp.</i></p><h2>July 25th</h2> <p>After a long break we finally started working on the actual assembly of parts - the mtr complex. Today Jonas ran a trial PCR on mtrB and mtrC part of the complex. GoTaq polymerase (Promega) was used, this is the polymerase we generally use for trials and colony PCRs as fidelity is not the main priority.</p><p><img src="https://static.igem.org/mediawiki/2013/c/cc/Gel-mtrBC.jpg" width="340" height="150" alt="mtrBCtrial" align="left">This is the gel from the PCR. Temperatures ranged 46-71˚C. It appears that the primers are sufficiently specific at even low temperatures. 1% Agarase, SYBR-Safe(Invitrogen), [http://www.york-bio.com/QStep4.jpg Q-Step4 ladder] </p> <h2>July 26th</h2> <p>Trial PCR again, this time for three synthetic DNA fragments - gBlocks (IDT), these fragments are highlighted below on the diagram as well as the overall view of our mtr gene. <img src="https://static.igem.org/mediawiki/2013/8/81/MtrABC.jpg" width="680" height="60" alt="mtrABC"></p> |
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<div id="drylabcontent"><p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Morbi in sem sodales, viverra nunc id, interdum tortor. Sed urna augue, dictum eget justo ut, dictum elementum massa. Nunc eu metus nunc. Aenean tempor sit amet quam accumsan vulputate. Curabitur nec tempus quam, quis fermentum leo. In laoreet venenatis arcu, sit amet elementum leo dignissim ut. Aenean id elementum nulla. Ut velit neque, lobortis id mollis quis, luctus sed tortor. Nam aliquam vitae orci et pharetra. Nunc ut metus in orci venenatis fermentum. Suspendisse placerat est purus, sagittis vehicula elit fermentum vitae. Aenean eleifend, odio sit amet semper dictum, dolor tortor feugiat nisl, ac ornare enim nisi sit amet purus. Cum sociis natoque penatibus et magnis dis parturient montes, nascetur ridiculus mus. Pellentesque sed turpis pretium, feugiat eros sit amet, consequat ligula. </p></div> | <div id="drylabcontent"><p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Morbi in sem sodales, viverra nunc id, interdum tortor. Sed urna augue, dictum eget justo ut, dictum elementum massa. Nunc eu metus nunc. Aenean tempor sit amet quam accumsan vulputate. Curabitur nec tempus quam, quis fermentum leo. In laoreet venenatis arcu, sit amet elementum leo dignissim ut. Aenean id elementum nulla. Ut velit neque, lobortis id mollis quis, luctus sed tortor. Nam aliquam vitae orci et pharetra. Nunc ut metus in orci venenatis fermentum. Suspendisse placerat est purus, sagittis vehicula elit fermentum vitae. Aenean eleifend, odio sit amet semper dictum, dolor tortor feugiat nisl, ac ornare enim nisi sit amet purus. Cum sociis natoque penatibus et magnis dis parturient montes, nascetur ridiculus mus. Pellentesque sed turpis pretium, feugiat eros sit amet, consequat ligula. </p></div> | ||
Revision as of 10:34, 26 September 2013
Overview
My part of the team - Gintare, Evaldas and Jonas - were responsible for cloning of the mtr complex, gold sensing device and preparing peptides for expression.
July 7th
Making of chemically competent DH5α cells. Protocol can be found in our protocols section.
July 10th
Purification of genomic DNA from Shewanella oneidensis and Salmonella sp.
July 25th
After a long break we finally started working on the actual assembly of parts - the mtr complex. Today Jonas ran a trial PCR on mtrB and mtrC part of the complex. GoTaq polymerase (Promega) was used, this is the polymerase we generally use for trials and colony PCRs as fidelity is not the main priority.
This is the gel from the PCR. Temperatures ranged 46-71˚C. It appears that the primers are sufficiently specific at even low temperatures. 1% Agarase, SYBR-Safe(Invitrogen), [http://www.york-bio.com/QStep4.jpg Q-Step4 ladder]
July 26th
Trial PCR again, this time for three synthetic DNA fragments - gBlocks (IDT), these fragments are highlighted below on the diagram as well as the overall view of our mtr gene. 
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Morbi in sem sodales, viverra nunc id, interdum tortor. Sed urna augue, dictum eget justo ut, dictum elementum massa. Nunc eu metus nunc. Aenean tempor sit amet quam accumsan vulputate. Curabitur nec tempus quam, quis fermentum leo. In laoreet venenatis arcu, sit amet elementum leo dignissim ut. Aenean id elementum nulla. Ut velit neque, lobortis id mollis quis, luctus sed tortor. Nam aliquam vitae orci et pharetra. Nunc ut metus in orci venenatis fermentum. Suspendisse placerat est purus, sagittis vehicula elit fermentum vitae. Aenean eleifend, odio sit amet semper dictum, dolor tortor feugiat nisl, ac ornare enim nisi sit amet purus. Cum sociis natoque penatibus et magnis dis parturient montes, nascetur ridiculus mus. Pellentesque sed turpis pretium, feugiat eros sit amet, consequat ligula.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Morbi in sem sodales, viverra nunc id, interdum tortor. Sed urna augue, dictum eget justo ut, dictum elementum massa. Nunc eu metus nunc. Aenean tempor sit amet quam accumsan vulputate. Curabitur nec tempus quam, quis fermentum leo. In laoreet venenatis arcu, sit amet elementum leo dignissim ut. Aenean id elementum nulla. Ut velit neque, lobortis id mollis quis, luctus sed tortor. Nam aliquam vitae orci et pharetra. Nunc ut metus in orci venenatis fermentum. Suspendisse placerat est purus, sagittis vehicula elit fermentum vitae. Aenean eleifend, odio sit amet semper dictum, dolor tortor feugiat nisl, ac ornare enim nisi sit amet purus. Cum sociis natoque penatibus et magnis dis parturient montes, nascetur ridiculus mus. Pellentesque sed turpis pretium, feugiat eros sit amet, consequat ligula.
The University of York
The University of York is Lorem ipsum dolor sit amet, consectetur adipiscing elit. Morbi in sem sodales, viverra nunc id, interdum tortor. Sed urna augue, dictum eget justo ut, dictum elementum massa. Nunc eu metus nunc. Aenean tempor sit amet quam accumsan vulputate. Curabitur nec tempus quam, quis fermentum leo. In laoreet venenatis arcu, sit amet elementum leo dignissim ut. Aenean id elementum nulla. Ut velit neque, lobortis id mollis quis, luctus sed tortor. Nam aliquam vitae orci et pharetra. Nunc ut metus in orci venenatis fermentum. Suspendisse placerat est purus, sagittis vehicula elit fermentum vitae. Aenean eleifend, odio sit amet semper dictum, dolor tortor feugiat nisl, ac ornare enim nisi sit amet purus. Cum sociis natoque penatibus et magnis dis parturient montes, nascetur ridiculus mus. Pellentesque sed turpis pretium, feugiat eros sit amet, consequat ligula.
