Team:CityU HK/notebook/lablog/project1

From 2014.igem.org

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                         <li>
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                             <a href="#">June<span class="st-arrow">Open or Close</span></a>
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                             <a href="#">July<span class="st-arrow">Open or Close</span></a>
                             <div class="st-content">
                             <div class="st-content">
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                                 <left><h1><b>Week 3 (13/7~19/7)</b></h1></left>
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                                 <h2 class="title"><b>Week 3 (13/7~19/7)</b></h2>
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<p>Primer design for leaderless tesA for Fit-coli (‘tesA) & New biobrick (‘tesA BB)
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                                <p><b>-</b> Primer design for leaderless tesA for Fit-coli (‘tesA) & New biobrick (‘tesA BB)</p>
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</p>
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                                <h2 class="title"><b>Week 4 (20/7~26/7)</b></h2>
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                                 <p>A small river named Duden flows by their place and supplies it with the necessary regelialia. It is a paradisematic country, in which roasted parts of sentences fly into your mouth.</p>
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                                 <p><b>-</b> PCR of ‘tesA and ‘tesA BB</p>
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                                 <img src="images/7.jpg" alt="image07"/>
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                                <p><b>-</b> PCR purification of ‘tesA and ‘tesA BB</p>
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                                 <img src="images/8.jpg" alt="image08"/>
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                                 <p><b>-</b> Prepare LB agar plates(‘tesA BB)</p>
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                                 <img src="images/9.jpg" alt="image09"/>
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                                <h2 class="title"><b>Week 5 (27/7~31/7)</b></h2>
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                                 <img src="images/10.jpg" alt="image10"/>
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                                 <p><b>-</b> Digestion of ‘tesA BB and pSB1C3 plasmid with EcoRI & PstI restriction enzymes</p>
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                                 <img src="images/11.jpg" alt="image11"/>
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                                 <p><b>-</b> PCR purification for digested ‘tesA BB</p>
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                                 <img src="images/12.jpg" alt="image12"/>
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                                 <p><b>-</b> Sub-cloning of ‘tesA BB into pSB1C3 plasmid</p>
 +
                                 <p><b>-</b> Transformation of ‘tesA BB into W3110 E.coli</p>
 +
                                 <p><b>-</b> Pick colonies of the transformed E. coli with ‘tesA BB</p>
                             </div>
                             </div>
                         </li>
                         </li>
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                         <li>
                         <li>
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                             <a href="#">Illustrations<span class="st-arrow">Open or Close</span></a>
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                             <a href="#">Early August<span class="st-arrow">Open or Close</span></a>
                             <div class="st-content">
                             <div class="st-content">
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                                 <p>O my friend - but it is too much for my strength - I sink under the weight of the splendour of these visions!</p>
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                                <h2 class="title"><b>Week 1 (1/8~2/8)</b></h2>
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                                 <p>A wonderful serenity has taken possession of my entire soul, like these sweet mornings of spring which I enjoy with my whole heart.</p>
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                                 <p><b>-</b> Cell lysis of picked E. coli with ‘tesA BB</p>
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                                 <p>I am alone, and feel the charm of existence in this spot, which was created for the bliss of souls like mine.</p>
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                                <p><b>-</b> Colonies PCR of ‘tesA BB by using VF2 & VR primer</p>
 +
                                <p><b>-</b> Send the ‘tesA BB plasmid for sequencing</p>
 +
 
 +
                                <h2 class="title"><b>Week 2 (3/8~9/8)</b></h2>
 +
                                <p><b>-</b> 2nd PCR of ‘tesA <br>∵ The first PCR product of ‘tesA was degraded over a long period of storage time</p>
 +
                                 <p><b>-</b> PCR purification of ‘tesA PCR product</p>
 +
                                <p><b>-</b> 1st Digestion of ‘tesA by using XbaI & PstI restriction enzyme<br>-> After running gel, no fragment showed in pilot ( other plasmid with the Xbal & Pstl respectively) , could not determine whether the digestion was success or not</p>
 +
                                 <p><b>-</b> 2nd digestion of ‘tesA by using XbaI & PstI restriction enzyme with adding one more control pilot that is only plasmid with no enzyme<br>-> Still no fragment presented in pilot, we wonder whether the pilot plasmid or the restriction enzyme had problem</p>
 +
                                <p><b>-</b> 3rd digestion of lac promoter in pSB1C3 plasmid & pbad promoter in pSB1C3 plasmid by using EcoRI, XbaI, SpeI & PstI restriction enzyme<br>-> All of the restriction enzyme worked normally, but the pbad promoter in pSB1C3 plasmid was degraded</p>
 +
                                <p><b>-</b> 4th digestion of ‘tesA by using XbaI & PstI restriction enzyme</p>
 +
                                <p><b>-</b> Sub-cloning of‘tesA into rbs in pSB1C3 plasmid (Part: BBa_B0030)</p>
 +
                               
                             </div>
                             </div>
                         </li>
                         </li>
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                         <li>
                         <li>
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                             <a href="#">Miscancellous<span class="st-arrow">Open or Close</span></a>
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                             <a href="#">Late August<span class="st-arrow">Open or Close</span></a>
                             <div class="st-content">
                             <div class="st-content">
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                                 <p>The bedding was hardly able to cover it and seemed ready to slide off any moment. His many legs, pitifully thin compared with the size of the rest of him, waved about helplessly as he looked. "What's happened to me?"</p>
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                                <h2 class="title"><b>Week 3 (10/8~16/8)</b></h2>
-
                                 <p>He thought. It wasn't a dream. His room, a proper human room although a little too small, lay peacefully between its four familiar walls.</p>
+
                                 <p><b>-</b> 1st Transformation of ‘tesA with rbs (BBa_B0030) into W3110 E.coli</p>
 +
                                <p><b>-</b> Pick colonies of the transformed E. coli with ‘tesA & rbs (BBa_B0030)</p>
 +
                                <p><b>-</b> Send the‘tesA with rbs (BBa_B0030) plasmid for sequencing<br>∵ The sequencing result showed the coding error of the rbs<br> -> Used BBa_B0034 rbs in pSB1C3 plasmid to replace BBa_B0030 rbs for 2nd cloning </p>
 +
                               
 +
 
 +
                                <h2 class="title"><b>Week 4 (17/8~23/8)</b></h2>
 +
                                <p><b>-</b> 3rd PCR of tesA’ <br>∵ The 2nd PCR product of ‘tesA was used up</p>
 +
                                <p><b>-</b> PCR purification of ‘tesA PCR product</p>
 +
                                <p><b>-</b> 5th digestion of ‘tesA by using XbaI and PstI restriction enzyme</p>
 +
                                <p><b>-</b> Sub-cloning of ‘tesA into rbs in pSB1A2 plasmid (Part: pSB1A2- BBa_B0034)</p>
 +
                                <p><b>-</b> Miniprep of pbad promoter (Part: pSB1C3-BBa_I13453)</p>
 +
                                <p><b>-</b> 2nd tramsformation of ‘tesA with rbs (BBa_B0034) into W3110 E.coli</p>
 +
                                <p><b>-</b> 1st digestion of pbad promoter (BBa_I13453) by using SpeI & PstI restriction enzyme</p>
 +
 
 +
                                <h2 class="title"><b>Week 5 (24/8~30/8)</b></h2>
 +
                                <p><b>-</b> Pick colonies of the transformed E. coli with ‘tesA & rbs (BBa_B0034)</p>
 +
                                 <p><b>-</b> Cell lysis of the picked E. coli with ‘tesA & rbs (BBa_B0034)</p>
 +
                                <p><b>-</b> Colonies PCR of ‘tesA & rbs (BBa_B0034) by using VF2 & VR primer</p>
 +
                                <p><b>-</b> Send the‘tesA with rbs (BBA-B0034) plasmid for sequencing</p>
 +
                                <p><b>-</b> Miniprep of ‘tesA with rbs (BBa_B0034)</p>
 +
                                <p><b>-</b> 1st digestion of ‘tesA with rbs (BBa_B0034) by using XbaI & PstI restriction enzyme</p>
 +
                                <p><b>-</b> Gel purification of digested ‘tesA with rbs (BBa_B0034)</p>
 +
                                <p><b>-</b> PCR purification of pbad promoter (BBa_I13453) digested in week 4</p>
                             </div>
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Latest revision as of 15:08, 11 September 2014

Bootstrap 101 Template



TesA module's lablog

  • JulyOpen or Close

    Week 3 (13/7~19/7)

    - Primer design for leaderless tesA for Fit-coli (‘tesA) & New biobrick (‘tesA BB)

    Week 4 (20/7~26/7)

    - PCR of ‘tesA and ‘tesA BB

    - PCR purification of ‘tesA and ‘tesA BB

    - Prepare LB agar plates(‘tesA BB)

    Week 5 (27/7~31/7)

    - Digestion of ‘tesA BB and pSB1C3 plasmid with EcoRI & PstI restriction enzymes

    - PCR purification for digested ‘tesA BB

    - Sub-cloning of ‘tesA BB into pSB1C3 plasmid

    - Transformation of ‘tesA BB into W3110 E.coli

    - Pick colonies of the transformed E. coli with ‘tesA BB

  • Early AugustOpen or Close

    Week 1 (1/8~2/8)

    - Cell lysis of picked E. coli with ‘tesA BB

    - Colonies PCR of ‘tesA BB by using VF2 & VR primer

    - Send the ‘tesA BB plasmid for sequencing

    Week 2 (3/8~9/8)

    - 2nd PCR of ‘tesA
    ∵ The first PCR product of ‘tesA was degraded over a long period of storage time

    - PCR purification of ‘tesA PCR product

    - 1st Digestion of ‘tesA by using XbaI & PstI restriction enzyme
    -> After running gel, no fragment showed in pilot ( other plasmid with the Xbal & Pstl respectively) , could not determine whether the digestion was success or not

    - 2nd digestion of ‘tesA by using XbaI & PstI restriction enzyme with adding one more control pilot that is only plasmid with no enzyme
    -> Still no fragment presented in pilot, we wonder whether the pilot plasmid or the restriction enzyme had problem

    - 3rd digestion of lac promoter in pSB1C3 plasmid & pbad promoter in pSB1C3 plasmid by using EcoRI, XbaI, SpeI & PstI restriction enzyme
    -> All of the restriction enzyme worked normally, but the pbad promoter in pSB1C3 plasmid was degraded

    - 4th digestion of ‘tesA by using XbaI & PstI restriction enzyme

    - Sub-cloning of‘tesA into rbs in pSB1C3 plasmid (Part: BBa_B0030)

  • Late AugustOpen or Close

    Week 3 (10/8~16/8)

    - 1st Transformation of ‘tesA with rbs (BBa_B0030) into W3110 E.coli

    - Pick colonies of the transformed E. coli with ‘tesA & rbs (BBa_B0030)

    - Send the‘tesA with rbs (BBa_B0030) plasmid for sequencing
    ∵ The sequencing result showed the coding error of the rbs
    -> Used BBa_B0034 rbs in pSB1C3 plasmid to replace BBa_B0030 rbs for 2nd cloning

    Week 4 (17/8~23/8)

    - 3rd PCR of tesA’
    ∵ The 2nd PCR product of ‘tesA was used up

    - PCR purification of ‘tesA PCR product

    - 5th digestion of ‘tesA by using XbaI and PstI restriction enzyme

    - Sub-cloning of ‘tesA into rbs in pSB1A2 plasmid (Part: pSB1A2- BBa_B0034)

    - Miniprep of pbad promoter (Part: pSB1C3-BBa_I13453)

    - 2nd tramsformation of ‘tesA with rbs (BBa_B0034) into W3110 E.coli

    - 1st digestion of pbad promoter (BBa_I13453) by using SpeI & PstI restriction enzyme

    Week 5 (24/8~30/8)

    - Pick colonies of the transformed E. coli with ‘tesA & rbs (BBa_B0034)

    - Cell lysis of the picked E. coli with ‘tesA & rbs (BBa_B0034)

    - Colonies PCR of ‘tesA & rbs (BBa_B0034) by using VF2 & VR primer

    - Send the‘tesA with rbs (BBA-B0034) plasmid for sequencing

    - Miniprep of ‘tesA with rbs (BBa_B0034)

    - 1st digestion of ‘tesA with rbs (BBa_B0034) by using XbaI & PstI restriction enzyme

    - Gel purification of digested ‘tesA with rbs (BBa_B0034)

    - PCR purification of pbad promoter (BBa_I13453) digested in week 4




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