Team:Fatih Turkey/Notebook8

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            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook">1</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook2">2</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook3">3</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook4">4</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook5">5</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook6">6</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook7">7</a></li>
            <li class="active"><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook8">8</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook9">9</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook10">10</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook11">11</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook12">12</a></li>
            <li><a href="https://2011.igem.org/Team:Fatih_Turkey/Notebook13">13</a></li>
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            <h4><strong>04.09.11</strong></h4><br>

1.       For each815, 815 gex, 825, 825 gex, 826, 926, 915, 915 gex, 925, 925 gex two colonies had been chosen and all were found wrong; thus this procedure is replied.<br>

2.       The parts above are isolated from their LBs.<br>

3.       Same parts are then digested with Xba1 and Pst1.<br>

4.       Same parts are loaded into gel.<br>

a.       For 915 and 915 gex, destinationplasmids were foundwrong; but genes were correct.<br>

b.      We were not so sure for destination plasmid of 826; but its gene were also correct.<br><br>


<a href="https://static.igem.org/mediawiki/2011/2/2a/04.09.11.png">https://static.igem.org/mediawiki/2011/2/2a/04.09.11.png</a><br>

<a href="https://static.igem.org/mediawiki/2011/4/45/04%2C09%2C2011.pdf">https://static.igem.org/mediawiki/2011/4/45/04%2C09%2C2011.pdf</a><br><br>

<h4><strong>05.09.11</strong></h4><br>

<a href="https://static.igem.org/mediawiki/2011/7/7a/05.09.11_-_II.png">https://static.igem.org/mediawiki/2011/7/7a/05.09.11_-_II.png</a><br>


<a href="https://static.igem.org/mediawiki/2011/5/56/05%2C09%2C11_DO%C4%9ERUSU.pdf">https://static.igem.org/mediawiki/2011/5/56/05%2C09%2C11_DO%C4%9ERUSU.pdf</a><br>


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<h4><strong>06.09.11</strong></h4><br>

1.       815 ep, 815 xp, 915 ep, 915 xp, 915 gexxp, 515 xparedigested-ligated. (Note that transformation is not performed because of absence of prepared compotent cell.)<br>

2.       545, 504-GFP and 915 aretransformated.<br>

3.       Each for 596, 596-b, 546, 536; two colonies are chosen. LBs for these parts are prepared. (8 LBs)<br>

4.       For 815, 815 gex, 915 and 915 gex; four additional LBs are prepared in order to gain more copies of them.<br>

5.       An antibiotic free LB is prepared to make compotent cell next day. One compotent cell in the stock is transferred into it.<br>

<a href="https://static.igem.org/mediawiki/2011/b/b1/06%2C09%2C2011.pdf">https://static.igem.org/mediawiki/2011/b/b1/06%2C09%2C2011.pdf</a><strong> </strong><br>

<a href="https://static.igem.org/mediawiki/2011/7/7a/05.09.11_-_II.png"></a><br>

<a href="https://static.igem.org/mediawiki/2011/4/4d/916_e%2Cp.pdf">https://static.igem.org/mediawiki/2011/4/4d/916_e%2Cp.pdf</a><br><br>


<h4><strong>07.09.11</strong></h4><br>

1.    We made  plasmid  isolation to 536H,546H,596B_G,596B_H,596H , j04500 but unfortunatelly  we fixed 596 G and 546 G in experiment  time .With xba1 and pst1:<br>

536H   &gt;&gt;&gt;&gt;                           13.1                                              29.4<br>

546H &gt;&gt;&gt;&gt;&gt;                            7.4                                               35.1<br>

596B_G &gt;&gt;&gt;&gt;                             15.1                                           27.4<br>

596B_H &gt;&gt;&gt;&gt;                       12.5                                                30<br>

596G+546&gt;&gt;&gt;&gt;                     11.6                                                  30.9<br>

596 H &gt;&gt;&gt;&gt;                             16.1                                                26.4<br>

With ecor1 and spe1:<br>

J04500_2(double dig.for up) &gt;&gt;&gt;&gt;         6.6                                                  35.9<br>

With e.cor1:<br>

J04500_2(single)&gt;&gt;&gt;&gt;     6.6                                                                       36.9<br>

J04500_2(undigest)&gt;&gt;&gt;&gt;   6.6                                                               37.9(WE FORGOT  THİS!!!    So we didnt  made electrophoresis j04500 _2 single and undigest and didnt put all of  j04500 _2(double dig.for up  )  to gel.<br>

2.then made them electrophoresis.<br>

3. 815 ,815 gex,545,916,j04450,can are  put liquid culture from glyserol stock. We choose  2 colonies from 915 gx xp   and also 2 colonies  from 504 gfp then put all of them 10 ml liquid culture.<br>

4.Top 10,DH 5alfa ,PET 28 a in BL21,BL 21 are put 10 ml liquid culture.<br><br>

<a href="https://static.igem.org/mediawiki/2011/1/1b/07%2C09%2C2011.confirmed.pdf">https://static.igem.org/mediawiki/2011/1/1b/07%2C09%2C2011.confirmed.pdf</a><br>

<a href="https://static.igem.org/mediawiki/2011/e/e5/07%2C09%2C2011.pdf">https://static.igem.org/mediawiki/2011/e/e5/07%2C09%2C2011.pdf</a><strong> </strong><br>


<a href="https://static.igem.org/mediawiki/2011/f/fe/07.09.11.png">https://static.igem.org/mediawiki/2011/f/fe/07.09.11.png</a><br><br><br>

<h4><strong>08.09.11</strong></h4><br><br>

1.       For each of 915 gexxpand 504-gfp, two colonies were incubated in 10 ml LB.<br>

2.       916, 900, 915 gexxp, 504-gfp, 545, 500, 815, 815 gex are isolated.<br>

3.       The parts above are digested with Xba1 and Pst1 except 545, 500.The separts are remainedun digested; because both of them had been confirmed.<br>

4.       Digested parts are loaded into gel.<br>

a.       All parts except 900 are found wrong.<br>


<a href="https://static.igem.org/mediawiki/2011/2/2b/08.09.11.png">https://static.igem.org/mediawiki/2011/2/2b/08.09.11.png</a><br>

<a href="https://static.igem.org/mediawiki/2011/e/e2/08%2C09%2C2011.pdf">https://static.igem.org/mediawiki/2011/e/e2/08%2C09%2C2011.pdf</a><br>
<br><br>

<h4><strong>09.09.11</strong></h4><br><br>

1.       Toligate 816, 825, 826, 916, 925, 926, 536 and 546; upstream, downstream and destination plasmids are digested with  appropiate enzymes.<br>

2.       Digested solutions are loaded into gel and all of them are confirmed.<br>

3.       For each of 815 and 915 parts in the stock; LB solutions are prepared.<br>

<a href="https://static.igem.org/mediawiki/2011/3/36/09.09.11.png">https://static.igem.org/mediawiki/2011/3/36/09.09.11.png</a><br><br>

<h4><strong>10.09.11</strong></h4><br><br>

1.       Because of absence of 800, digested solutions that prepared for the ligation of 816, 825 and 826 are <strong>not</strong>ligated.<br>

2.       536, 546, 916, 925 and 926 are ligated.<br>

3.       916, 925 and 926 transformated twice. (We have two plates each of these parts.) 536 and 546 are transformated into only one plate.<br>

4.       LBs that were prepared yesterday are prepared again. (Isolation could not take place today.)<br><br>


<h4><strong>11.09.11</strong></h4>
<a href="https://static.igem.org/mediawiki/2011/a/a5/11%2C09%2C2011_2.pdf">https://static.igem.org/mediawiki/2011/a/a5/11%2C09%2C2011_2.pdf</a><br>

<a href="https://static.igem.org/mediawiki/2011/0/0d/11%2C09%2C2011.pdf">https://static.igem.org/mediawiki/2011/0/0d/11%2C09%2C2011.pdf</a><br><br>
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