Oscarliu117 (Talk | contribs) |
Huiyee11186 (Talk | contribs) |
||
(27 intermediate revisions by 4 users not shown) | |||
Line 11: | Line 11: | ||
<!--Page_Content--> | <!--Page_Content--> | ||
<div class="container content"> | <div class="container content"> | ||
+ | <div class="headstyle"> | ||
+ | <h1 class="head">Notebook</h1> | ||
+ | </div> | ||
+ | <div class="righttitle"> | ||
+ | <h6 class="subtitle">Of Blood, Sweat and Tears</h6> | ||
+ | </div> | ||
<div class="navbar-example"> | <div class="navbar-example"> | ||
<div class="row"> | <div class="row"> | ||
+ | <div class="col-2 side"> | ||
+ | <div id="sidelist" class="list-group"> | ||
+ | <a class="list-group-item list-group-item-action" href="#Pasr_and_Pgada">P<sub>asr</sub> and P<sub>gadA</sub></a> | ||
+ | <a class="list-group-item list-group-item-action" href="#pH_sensor_test">pH Sensor Test</a> | ||
+ | <a class="list-group-item list-group-item-action" href="#CA">CA</a> | ||
+ | <a class="list-group-item list-group-item-action" href="#RbcL">RbcL</a> | ||
+ | <a class="list-group-item list-group-item-action" href="#RbcXS">RbcXS</a> | ||
+ | <a class="list-group-item list-group-item-action" href="#PRK">PRK</a> | ||
+ | <a class="list-group-item list-group-item-action" href="#Whole_Construction">Whole Construction</a> | ||
+ | <a class="list-group-item list-group-item-action" href="#Total_Solution">Total Solution</a> | ||
+ | <a class="list-group-item list-group-item-action" href="#"><i class="fa fa-arrow-up fa-1x" | ||
+ | aria-hidden="true"></i> | ||
+ | </a> | ||
+ | </div> | ||
+ | </div> | ||
<div class="col-10"> | <div class="col-10"> | ||
<div data-spy="scroll" data-target="#sidelist" data-offset="0" class="scrollspy-example"> | <div data-spy="scroll" data-target="#sidelist" data-offset="0" class="scrollspy-example"> | ||
+ | <div id="Pasr_and_Pgada"> | ||
+ | <h3>Construction of P<sub>asr</sub> and P<sub>gadA</sub></h3></br> | ||
− | + | <p class="pcontent">4/26~4/27</br> </p> | |
+ | <ul> | ||
+ | <li>PCR P<sub>asr</sub> and P<sub>gadA</sub> from <i>E. coli</i> MG1655 chromosome. Confirmed and extracted | ||
+ | products by DNA gel. </li> | ||
+ | </ul></br> | ||
− | + | <p class="pcontent">4/28</br> </p> | |
+ | <ul> | ||
+ | <li>Digested the pSB1C3-GFP plasmid. Ligase the plasmid with P<sub>asr</sub> / P<sub>gadA</sub> and | ||
+ | RiboJ by PCR. Confirmed by DNA gel electrophoresis. Transformed the gene in DH5 | ||
+ | alpha and spread the bacteria on LB plates.</li> | ||
+ | </ul></br> | ||
− | + | <p class="pcontent">4/30~5/1</br> </p> | |
− | + | <ul> | |
− | + | <li>Confirmed the transformation by colony PCR ,enzyme digestion and DNA gel | |
− | + | electrophoresis. Selected the correct colonies for sequencing and cultured them | |
− | + | on LB plates.</li> | |
+ | </ul></br> | ||
− | + | <p class="pcontent">5/9</br> </p> | |
− | + | <ul> | |
− | + | <li>Store the correct colonies in glycerol in -80°C.</li> | |
− | + | </ul></br> | |
− | + | ||
− | + | ||
− | + | <p class="pcontent">6/1</br> </p> | |
− | + | <ul> | |
− | + | <li>Transformed each plasmid into BL21(DE3)</li> | |
− | + | </ul></br> | |
− | + | </div> | |
− | + | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | + | <div id="pH_sensor_test"> | |
− | + | <h3>The pH sensor test</h3></br> | |
− | + | ||
− | + | ||
− | + | ||
− | + | <p class="pcontent">5/9</br> </p> | |
− | + | <ul> | |
− | + | <li>Prepared the M9 medium in different pH value.</li> | |
− | + | </ul></br> | |
− | + | <p class="pcontent">5/11</br> </p> | |
− | + | <ul> | |
− | + | <li>Pre-cultured the DH5 alpha-pSB1C3 -P<sub>asr</sub>-sfGFP strain.</li> | |
− | + | </ul></br> | |
− | + | <p class="pcontent">5/12</br> </p> | |
− | + | <ul> | |
− | + | <li>Cultured the DH5 alpha-pSB1C3 -P<sub>asr</sub>-sfGFP strain in M9 medium in different pH | |
− | + | value. | |
− | + | Measured the O.D. 600 value and fluorescence every hour.</li> | |
+ | </ul></br> | ||
− | + | <p class="pcontent">5/18</br> </p> | |
− | + | <ul> | |
− | + | <li>Pre-cultured DH5 alpha-pSB1C3 –P<sub>gadA</sub>-RiboJ-sfGFP strain.</li> | |
− | + | <li>Prepared the M9 medium in different pH value.</li> | |
− | + | </ul></br> | |
− | + | <p class="pcontent">5/19</br> </p> | |
− | + | <ul> | |
− | + | <li>Cultured the DH5 alpha-pSB1C3 –P<sub>gadA</sub>-RiboJ-sfGFP strain in M9 medium in different | |
− | + | pH | |
− | + | value. Measured the O.D. 600 value and fluorescence every hour.</li> | |
+ | </ul></br> | ||
− | + | <p class="pcontent">6/2~6/3</br> </p> | |
− | + | <ul> | |
− | + | <li>Repeated the experiment on 5/11~5/12</li> | |
− | + | </ul></br> | |
− | + | <p class="pcontent">7/29</br> </p> | |
− | + | <ul> | |
− | + | <li>Pre-culture the BL21(DE3)-pSB1C3 –P<sub>asr</sub>-sfGFP strain</li> | |
− | + | </ul></br> | |
− | + | <p class="pcontent">7/30</br> </p> | |
− | + | <ul> | |
− | + | <li>Cultured the BL21(DE3)-pSB1C3 –P<sub>asr</sub> -sfGFP strain on 96 well plate in different pH | |
− | + | value. Measure the fluorescence every 5 minutes in 30 minutes.</li> | |
− | + | </ul></br> | |
− | + | <p class="pcontent">9/4</br> </p> | |
− | + | <ul> | |
− | + | <li>Pre-culture the BL21(DE3)-pSB1C3 –P<sub>gadA</sub>-RiboJ-sfGFP strain</li> | |
− | + | </ul></br> | |
− | + | <p class="pcontent">9/5</br> </p> | |
− | + | <ul> | |
− | + | <li>Cultured the BL21(DE3)-pSB1C3 –P<sub>gadA</sub>-RiboJ-sfGFP strain strain on 96 well plate in | |
− | + | different pH value. Measure the fluorescence every 5 minutes in 30 minutes.</li> | |
− | + | </ul></br> | |
+ | </div> | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | + | <div id="CA"> | |
− | + | <h3>Construction of CA</h3></br> | |
− | + | <p class="pcontent">7/2</br> </p> | |
− | + | <ul> | |
− | + | <li>PCR IDT products (RbcL, RbcXS, PRK, CA). Confirmed by DNA gel electrophoresis. | |
− | + | Extracted the gene from DNA gel. | |
− | + | ||
− | + | </li> | |
− | + | </ul></br> | |
− | + | <p class="pcontent">7/5</br> </p> | |
− | + | <ul> | |
− | + | <li>PCR the CA. Confirmed by DNA gel electrophoresis and extracted the gene from DNA | |
− | + | gel. | |
− | + | </li> | |
− | + | </ul></br> | |
− | + | ||
− | + | ||
− | + | <p class="pcontent"> | |
− | + | 7/6 | |
− | + | </br> </p> | |
− | + | <ul> | |
+ | <li>Digested and ligase the pSB1C3 plasmid and CA gene. | ||
+ | </li> | ||
+ | </ul></br> | ||
+ | <p class="pcontent">7/10~7/11</br> </p> | ||
+ | <ul> | ||
+ | <li>Repeated the PCR of CA, digestion, ligation.</li> | ||
+ | </ul></br> | ||
+ | <p class="pcontent">7/12</br> </p> | ||
+ | <ul> | ||
+ | <li>Transformed the pSB1C3-P<sub>T7</sub>-CA into DH5 alpha. </li> | ||
+ | </ul></br> | ||
+ | <p class="pcontent">7/19</br> </p> | ||
+ | <ul> | ||
+ | <li>Send the plasmid to sequencing</li> | ||
+ | </ul></br> | ||
+ | </div> | ||
− | |||
− | < | + | <div id="RbcL"> |
− | + | <h3>Construction of RbcL</h3></br> | |
− | < | + | |
− | + | ||
− | + | ||
− | |||
− | |||
− | |||
− | |||
− | + | <p class="pcontent">7/2</br> </p> | |
− | + | <ul> | |
− | + | <li>PCR IDT products (RbcL, RbcXS, PRK, CA). Confirmed by DNA gel electrophoresis.</li> | |
− | + | <li>Extracted the gene from DNA gel.</li> | |
+ | </ul></br> | ||
− | + | <p class="pcontent">7/11</br> </p> | |
− | + | <ul> | |
− | + | <li>PCR the RbcL with new primer.</li> | |
− | + | </ul></br> | |
+ | <p class="pcontent">7/20</br> </p> | ||
+ | <ul> | ||
+ | <li>PCR the RbcL with old and new primer.</li> | ||
+ | </ul></br> | ||
− | + | <p class="pcontent">7/21~24</br> </p> | |
− | + | <ul> | |
− | + | <li>Repeated the PCR of RbcL</li> | |
− | + | </ul></br> | |
− | + | ||
− | |||
− | |||
− | |||
− | |||
− | |||
+ | <p class="pcontent">8/1</br> </p> | ||
+ | <ul> | ||
+ | <li>Ligate the RbcL with P<sub>LacI</sub>.</li> | ||
+ | <li>Transformed the pSB1C3-P<sub>T7</sub>-RbcL and pSB1C3-P<sub>LacI</sub>-RbcL into DH5 alpha.</li> | ||
+ | </ul></br> | ||
+ | <p class="pcontent">8/3 </br> </p> | ||
+ | <ul> | ||
+ | <li>Confirmed the pSB1C3-P<sub>T7</sub>-RbcL construct by colony PCR.</li> | ||
+ | <li>Sent the gene to sequencing</li> | ||
+ | </ul></br> | ||
+ | </div> | ||
+ | <div id="RbcXS"> | ||
+ | <h3>Construction of RbcXS</h3></br> | ||
+ | <p class="pcontent">7/2</br> </p> | ||
+ | <ul> | ||
+ | <li>Prepared the M9 medium in different pH value.</li> | ||
+ | </ul></br> | ||
+ | <p class="pcontent">7/3~7/21</br> </p> | ||
+ | <ul> | ||
+ | <li>PCR IDT products (RbcL, RbcXS, PRK, CA). Confirmed by DNA gel electrophoresis.</li> | ||
+ | <li> Extracted the gene from DNA gel.</li> | ||
+ | </ul></br> | ||
+ | <p class="pcontent">7/23</br> </p> | ||
+ | <ul> | ||
+ | <li>PCR normal/mutant RbcXS with old and new primer.</li> | ||
+ | </ul></br> | ||
− | + | <p class="pcontent">7/24 </br> </p> | |
+ | <ul> | ||
+ | <li>Confirmed by DNA gel.</li> | ||
+ | </ul></br> | ||
+ | </div> | ||
− | |||
− | |||
− | |||
− | |||
+ | <div id="PRK"> | ||
+ | <h3>Construction of PRK</h3></br> | ||
− | + | <p class="pcontent">7/2</br> </p> | |
+ | <ul> | ||
+ | <li>PCR IDT products (RbcL, RbcXS, PRK, CA). Confirmed by DNA gel electrophoresis.</li> | ||
+ | <li> Extracted the gene from DNA gel.</li> | ||
+ | </ul></br> | ||
− | + | <p class="pcontent">7/3~8/2</br> </p> | |
+ | <ul> | ||
+ | <li>PCR PRK.</li> | ||
+ | </ul></br> | ||
− | <p class="pcontent"> | + | <p class="pcontent">8/1</br> </p> |
− | <ul> | + | <ul> |
− | <li> | + | <li>Ligated PRK with P<sub>LacI</sub> and P<sub>T7</sub>.</li> |
− | + | <li>Transformed into DH5 alpha. | |
+ | |||
+ | </li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">8/2</br> </p> | ||
+ | <ul> | ||
+ | <li>Confirmed by colony PCR. Selected the colony with correct length and sent to | ||
+ | sequencing.</li> | ||
+ | </ul></br> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | <div id="Whole_Construction"> | ||
+ | <h3>Whole construction</h3></br> | ||
+ | |||
+ | <p class="pcontent">8/8</br> </p> | ||
+ | <ul> | ||
+ | <li>Ligated P<sub>T7</sub>-CA with P<sub>LacI</sub>-PRK on pSB1C3</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">8/9</br> </p> | ||
+ | <ul> | ||
+ | <li>Ligated P<sub>T7</sub>-RbcL with P<sub>T7</sub>-RbcXS on pSB1C3</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">8/28</br> </p> | ||
+ | <ul> | ||
+ | <li>Inserted P<sub>T7</sub>-CA with P<sub>LacI</sub>-PRK into pSB3K3</li> | ||
+ | </ul></br> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <div id="Total_Solution"> | ||
+ | <h3>SDS PAGE, Functional test and Total solution</h3></br> | ||
+ | |||
+ | <p class="pcontent">7/25~7/30, 8/4~8/5, 8/11~8/12</br> </p> | ||
+ | <ul> | ||
+ | <li>Ran SDS-PAGE of CA, normal-RbcXS </li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">8/7, 8/9, 8/10</br> </p> | ||
+ | <ul> | ||
+ | <li> Ran SDS-PAGE of RbcL and PRK</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">8/16 </br> </p> | ||
+ | <ul> | ||
+ | <li>Ran the SDS-PAGE of RbcL, RbcXS and PRK</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">8/18~8/19</br> </p> | ||
+ | <ul> | ||
+ | <li>Activity test of CA</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">9/2</br> </p> | ||
+ | <ul> | ||
+ | <li> Ran SDS-PAGE of pSB1C3-P<sub>T7</sub>-RbcL-P<sub>T7</sub>-RbcXS and pSB3K3-P<sub>LacI</sub>-PRK-P<sub>T7</sub>-CA</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">9/3</br> </p> | ||
+ | <ul> | ||
+ | <li> Transformed pSB1C3-P<sub>T7</sub>-RbcL-P<sub>T7</sub>-RbcXS and pSB3K3-P<sub>LacI</sub>-PRK-P<sub>T7</sub>-CA into W3110</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">9/5~9/17</br> </p> | ||
+ | <ul> | ||
+ | <li> Total solution.</li> | ||
+ | </ul></br> | ||
+ | </div> | ||
Line 232: | Line 349: | ||
</div> | </div> | ||
</div> | </div> | ||
+ | |||
+ | |||
+ | <script> | ||
+ | $(document).ready(function () { | ||
+ | $(window).scroll(function () { | ||
+ | if ($(this).scrollTop() >= 500) { | ||
+ | var position = $("#sidelist").position(); | ||
+ | if (position == undefined) {} else { | ||
+ | $('#sidelist').css({ | ||
+ | "position": "fixed", | ||
+ | "top": "145px", | ||
+ | "margin-top": "0px" | ||
+ | }); | ||
+ | } | ||
+ | } else { | ||
+ | $('#sidelist').removeAttr('style'); | ||
+ | } | ||
+ | }); | ||
+ | $(function () { | ||
+ | $('i.fa-arrow-up').click(function () { | ||
+ | $('html, body').animate({ | ||
+ | scrollTop: 0 | ||
+ | }, 600); | ||
+ | return false; | ||
+ | }); | ||
+ | }); | ||
+ | }); | ||
+ | </script> | ||
+ | |||
</body> | </body> | ||
</html> | </html> | ||
{{NCKU_Tainan/footer}} | {{NCKU_Tainan/footer}} |
Latest revision as of 20:13, 17 October 2018