Team:CityU HK/notebook/lablog/project1

From 2014.igem.org

(Difference between revisions)
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                         <li>
                         <li>
-
                             <a href="#">August<span class="st-arrow">Open or Close</span></a>
+
                             <a href="#">Early August<span class="st-arrow">Open or Close</span></a>
                             <div class="st-content">
                             <div class="st-content">
                                 <h2 class="title"><b>Week 1 (1/8~2/8)</b></h2>
                                 <h2 class="title"><b>Week 1 (1/8~2/8)</b></h2>
Line 86: Line 86:
                                 <h2 class="title"><b>Week 2 (3/8~9/8)</b></h2>
                                 <h2 class="title"><b>Week 2 (3/8~9/8)</b></h2>
-
                                 <p>- 2¬nd PCR of ‘tesA <br>(∵ The first PCR product of ‘tesA was degraded over a long period of storage time)</p>
+
                                 <p>- 2nd PCR of ‘tesA <br>∵ The first PCR product of ‘tesA was degraded over a long period of storage time</p>
-
                                 <p>- PCR purification of ‘tesA and ‘tesA BB</p>
+
                                 <p>- PCR purification of ‘tesA PCR product</p>
-
                                 <p>- Prepare LB agar plates(‘tesA BB)</p>
+
                                 <p>- 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>- 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>- 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>- 4th digestion of ‘tesA by using XbaI & PstI restriction enzyme</p>
 +
                                <p>- Sub-cloning of‘tesA into rbs in pSB1C3 plasmid (Part: BBa_B0030)</p>
 +
 
                                 <h2 class="title"><b>Week 5 (27/7~31/7)</b></h2>
                                 <h2 class="title"><b>Week 5 (27/7~31/7)</b></h2>
                                 <p>- Digestion of ‘tesA BB and pSB1C3 plasmid with EcoRI & PstI restriction enzymes</p>
                                 <p>- Digestion of ‘tesA BB and pSB1C3 plasmid with EcoRI & PstI restriction enzymes</p>
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                         <li>
                         <li>
-
                             <a href="#">Miscancellous<span class="st-arrow">Open or Close</span></a>
+
                             <a href="#">Late August<span class="st-arrow">Open or Close</span></a>
                             <div class="st-content">
                             <div class="st-content">
-
                                 <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>
+
                                <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>- Cell lysis of picked E. coli with ‘tesA BB</p>
 +
                                <p>- Colonies PCR of ‘tesA BB by using VF2 & VR primer</p>
 +
                                <p>- Send the ‘tesA BB plasmid for sequencing</p>
 +
 
 +
                                <h2 class="title"><b>Week 2 (3/8~9/8)</b></h2>
 +
                                <p>- 2nd PCR of ‘tesA <br>∵ The first PCR product of ‘tesA was degraded over a long period of storage time</p>
 +
                                <p>- PCR purification of ‘tesA PCR product</p>
 +
                                <p>- 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>- 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>- 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>- 4th digestion of ‘tesA by using XbaI & PstI restriction enzyme</p>
 +
                                <p>- Sub-cloning of‘tesA into rbs in pSB1C3 plasmid (Part: BBa_B0030)</p>
 +
 
 +
                                <h2 class="title"><b>Week 5 (27/7~31/7)</b></h2>
 +
                                <p>- Digestion of ‘tesA BB and pSB1C3 plasmid with EcoRI & PstI restriction enzymes</p>
 +
                                 <p>- PCR purification for digested ‘tesA BB</p>
 +
                                <p>- Sub-cloning of ‘tesA BB into pSB1C3 plasmid</p>
 +
                                <p>- Transformation of ‘tesA BB into W3110 E.coli</p>
 +
                                <p>- Pick colonies of the transformed E. coli with ‘tesA BB</p>
                             </div>
                             </div>
                         </li>
                         </li>

Revision as of 14:43, 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)

    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

  • Late AugustOpen or Close

    Week 3 (10/8~16/8)

    - 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)

    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

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