Oscarliu117 (Talk | contribs) |
Oscarliu117 (Talk | contribs) |
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<h3>Construction of Pasr and Pgada</h3></br> | <h3>Construction of Pasr and Pgada</h3></br> | ||
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<h3>Construction of CA</h3></br> | <h3>Construction of CA</h3></br> | ||
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+ | <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> | ||
− | |||
− | <p class="pcontent"> | + | |
+ | |||
+ | |||
+ | |||
+ | <h3>Construction of PRK</h3></br> | ||
+ | |||
+ | <p class="pcontent">7/2</br> </p> | ||
<ul> | <ul> | ||
− | <li> | + | <li>PCR IDT products (RbcL, RbcXS, PRK, CA). Confirmed by DNA gel electrophoresis.</li> |
+ | <li> Extracted the gene from DNA gel.</li> | ||
</ul></br> | </ul></br> | ||
+ | <p class="pcontent">7/3~8/2</br> </p> | ||
+ | <ul> | ||
+ | <li>PCR PRK.</li> | ||
+ | </ul></br> | ||
+ | <p class="pcontent">8/1</br> </p> | ||
+ | <ul> | ||
+ | <li>Ligated PRK with PlacI and PT7.</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> | ||
− | |||
− | <p class="pcontent"> | + | |
+ | |||
+ | <h3>Whole construction</h3></br> | ||
+ | |||
+ | <p class="pcontent">8/8</br> </p> | ||
<ul> | <ul> | ||
− | <li> | + | <li>Ligated PT7-CA with PLacI-PRK on pSB1C3</li> |
</ul></br> | </ul></br> | ||
+ | |||
+ | <p class="pcontent">8/9</br> </p> | ||
+ | <ul> | ||
+ | <li>Ligated PT7-RbcL with PT7-RbcXS on pSB1C3</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">8/28</br> </p> | ||
+ | <ul> | ||
+ | <li>Inserted PT7-CA with PLacI-PRK into pSB3K3</li> | ||
+ | </ul></br> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <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-PT7-RbcL-PT7-RbcXS and pSB3K3-Plac-PRK-PT7-CA</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">9/3</br> </p> | ||
+ | <ul> | ||
+ | <li> Transformed pSB1C3-PT7-RbcL-PT7-RbcXS and pSB3K3-Plac-PRK-PT7-CA into W3110</li> | ||
+ | </ul></br> | ||
+ | |||
+ | <p class="pcontent">9/5~9/17</br> </p> | ||
+ | <ul> | ||
+ | <li> Total solution.</li> | ||
+ | </ul></br> | ||
+ | |||
Revision as of 05:34, 22 September 2018