Difference between revisions of "Team:SPSingapore/Notebook-Week-2"

 
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<div id='cssmenu'>
 
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<ul>
 
<ul>
−
  <li><a href='https://2015.igem.org/Team:SPSingapore/'><span>Home</span></a></li>
+
<li class = 'active'><a href='https://2015.igem.org/Team:SPSingapore/'><span>Home</span></a></li>
−
  <li><a href='https://2015.igem.org/Team:SPSingapore/Team'><span>Team</span></a></li>
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<li><a href='https://2015.igem.org/Team:SPSingapore/Team'><span>Team</span></a>
−
  <li><a href='https://2015.igem.org/Team:SPSingapore/Project'><span>Project</span></a></li>
+
<ul>
−
  <li><a href='https://2015.igem.org/Team:SPSingapore/Protocol'><span>Protocol</span></a></li>
+
<li><a href="https://2015.igem.org/Team:SPSingapore/Team">Overview</a></li>
−
  <li><a href='https://2015.igem.org/Team:SPSingapore/Parts'><span>Parts</span></a></li>
+
<li><a href="https://igem.org/Team.cgi?id=1804">Official Profile</a></li>
−
  <li class = 'active'><a href='https://2015.igem.org/Team:SPSingapore/Notebook'><span>Notebook</span></a></li>
+
        <li><a href="https://2015.igem.org/Team:SPSingapore/Team Bios">Team Bios</a></li>
−
  <li><a href='https://2015.igem.org/Team:SPSingapore/Practices'><span>Human Practices</span></a></li>
+
        <li><a href="https://2015.igem.org/Team:SPSingapore/Mentors">Mentors</a></li>
−
  <li class='last'><a href='https://2015.igem.org/Team:SPSingapore/Safety'><span>Safety</span></a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Attributions">Attributions</a></li>
 +
</ul>
 +
</li>
 +
<li><a href='https://2015.igem.org/Team:SPSingapore/Project'><span>Project</span></a>
 +
<ul>
 +
<li><a href='https://2015.igem.org/Team:SPSingapore/Project'>Overview</a></li>
 +
        <li><a href="https://2015.igem.org/Team:SPSingapore/Invasin">Invasin + Listerolysin</a></li>
 +
        <li><a href="https://2015.igem.org/Team:SPSingapore/ESAQS">esa Quorum Sensing</a></li>
 +
        <li><a href="https://2015.igem.org/Team:SPSingapore/Anaerobic Promoter">Anaerobic Promoter</a></li>
 +
        <li><a href="https://2015.igem.org/Team:SPSingapore/Parts">Parts</a></li>
 +
</ul>
 +
</li>
 +
<li><a href='https://2015.igem.org/Team:SPSingapore/Notebook'><span>Notebook</span></a>
 +
<ul>
 +
<li><a href="https://2015.igem.org/Team:SPSingapore/Protocol">Protocols</a></li>
 +
        <li><a href="https://2015.igem.org/Team:SPSingapore/Notebook">Entries</a></li>
 +
</ul>
 +
</li>
 +
<li><a href='https://2015.igem.org/Team:SPSingapore/Practices'><span>Human Practices</span></a>
 +
<ul>
 +
<li><a href="https://2015.igem.org/Team:SPSingapore/Practices">Overview</a></li>
 +
<li><a href="https://2015.igem.org/Team:SPSingapore/Workshop">Workshop</a></li>
 +
<li><a href="https://2015.igem.org/Team:SPSingapore/Workshop Materials">Workshop Materials</a></li>
 +
        <li><a href="https://2015.igem.org/Team:SPSingapore/Interview">Consultations</a></li>
 +
</ul>
 +
</li>
 +
<li><a href='https://2015.igem.org/Team:SPSingapore/Safety'><span>Safety</span></a></li>
 +
<li class='last'><a href='https://2015.igem.org/Team:SPSingapore/Medals'><span>Medals</span></a></li>
 
</ul>
 
</ul>
 
</div>
 
</div>
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<div id='sidemenu' style = "float:left">
 
<div id='sidemenu' style = "float:left">
 
<ul>
 
<ul>
 +
  <li><a href = "https://2015.igem.org/Team:SPSingapore/Notebook"><span>NOTEBOOK</span></a>
 +
  <ul><li class = 'last'>&nbsp;</li></ul></li>
 +
 
 +
  <li><a href = "https://2015.igem.org/Team:SPSingapore/Protocol"><span>PROTOCOL</span></a>
 +
  <ul><li class = 'last'>&nbsp;</li></ul></li>
 +
 
   <li class='last'><a href = "https://2015.igem.org/Team:SPSingapore/Notebook"><span>ENTRIES</span></a>
 
   <li class='last'><a href = "https://2015.igem.org/Team:SPSingapore/Notebook"><span>ENTRIES</span></a>
 
<ul>
 
<ul>
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         <li><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-15"><span>Week 15 (30/8 - 5/9)</span></a></li>
 
         <li><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-15"><span>Week 15 (30/8 - 5/9)</span></a></li>
 
         <li><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-16"><span>Week 16 (6/9 - 12/9)</span></a></li>
 
         <li><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-16"><span>Week 16 (6/9 - 12/9)</span></a></li>
−
         <li class = 'last'><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-17"><span>Week 17 (13/8 - 17/9)</span></a></li>
+
         <li class = 'last'><a href = "https://2015.igem.org/Team:SPSingapore/Notebook-Week-17"><span>Week 17 (13/9 - 18/9)</span></a></li>
 
       </ul>
 
       </ul>
 
   </li>
 
   </li>
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<table><tr><td>
 
<table><tr><td>
 
<div style = "border-top: 5px solid blue; width:750px; text-align:justify;margin-bottom: 20px; line-height:normal;">
 
<div style = "border-top: 5px solid blue; width:750px; text-align:justify;margin-bottom: 20px; line-height:normal;">
−
<h3 style ="margin-left:20px;">Week 2 (31/5 - 6/6)</h3>
+
<h3 style ="margin-left:20px;">Week 16 (6/9 - 12/9)</h3>
  
  
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<!----------------------- Entry Start ------------------------>
 
<!----------------------- Entry Start ------------------------>
 
<center>
 
<center>
−
<table style = "width:700px; border-collapse: collapse;">
+
<table style = "width:750px; border-collapse: collapse;">
  
−
<tr class = "tablenotebook"><td><p class = "paper">
+
<! ---------DAY ONE ------------------------------->
−
May 31
+
 
−
<br><br>
+
<tr class = "tablenotebook"><td><div class = "paper">
−
<strong>Xin Yi</strong>
+
&#9642; May 31
 
<br><br>
 
<br><br>
−
&bull; Sent EsaR clone 4 and INv-4 for sequencing.
+
<span class = "blue"> Invasin + Listerolysin</span>
 +
&emsp;&emsp; <span class = "black"> esa Quorum Sensing</span>
 
<br>
 
<br>
−
</p>
+
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Xin Yi &#9728;</font>
 +
<br><br>
 +
<div class = "divnbcontent">
 +
<span class = "nbcontent">
 +
Sent EsaR clone 4 and INv-4 for sequencing.
 +
</span>
 +
</div>
 +
 
 +
<br><br>
 +
 
 +
<span class = "blue"> Invasin + Listerolysin</span>
 +
&emsp;&emsp;
 +
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Yan Ting &#9728;</font>
 +
<br><br>
 +
<div class = "divnbcontent">
 +
<span class = "nbcontent">
 +
RE with XbaI and PstI
 +
<br>&emsp;&emsp;&emsp;&emsp; RE - 2ul
 +
<br>&emsp;&emsp;&emsp;&emsp; Buffer - 5ul
 +
<br>&emsp;&emsp;&emsp;&emsp; DNA (4x of miniprep=74.5ng/ul) - 13.4ul
 +
<br>&emsp;&emsp;&emsp;&emsp; H2O - 29.6ul
 +
<br>&emsp;&emsp;&emsp;&emsp; Incubate at 37degC for 2.5h (1130-1400)
 +
</span>
 +
<br><br>
 +
<span>
 +
&#9654; Results: Gel loaded 1kb ladder, uncut, RE digested. 100V for 1h.
 +
<br> Plasmid doesn't have XbaI site - RE digested DNA is a linear 6kb band.
 +
</span>
 +
 
 +
</div>
 +
 
 +
<br><br>
 +
 
 +
 
 +
</div>
 +
<div style = "border-top: 2px solid turquoise;"></div>
 
</td>
 
</td>
 
</tr>
 
</tr>
  
−
<tr class = "tablenotebook"><td><p class = "paper">
+
 
−
June 2
+
<!---------------next day ----------------------------------------->
 +
 
 +
<tr class = "tablenotebook"><td><div class = "paper">
 +
&#9642; June 2
 
<br><br>
 
<br><br>
−
<strong>Yi Han</strong>
+
<span class = "brown"> Anaerobic Promoter</span>
 +
&emsp;&emsp;
 +
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Xin Yi &#9728;</font>
 
<br><br>
 
<br><br>
−
&bull; YFP and GFP transformation results - no colonies for YFP<br>
+
<div class = "divnbcontent">
−
&bull; The GFP transformation repeated with 100ng of plasmid was sucessful.<br>
+
<span class = "nbcontent">
−
&bull; 4 colonies of gfp plasmid were inoculated in 3mL LB+amp and grown overnight.
+
YFP and GFP transformation results - no colonies for YFP
−
<br>
+
</span><br>
−
</p>
+
<span class = "nbcontent">
 +
The GFP transformation repeated with 100ng of plasmid was sucessful. 4 colonies of gfp plasmid were inoculated in 3mL LB+amp and grown overnight.
 +
</span>
 +
</div>
 +
 
 +
<br><br>
 +
 
 +
</div>
 +
<div style = "border-top: 2px solid turquoise;"></div>
 
</td>
 
</td>
 
</tr>
 
</tr>
  
−
<tr class = "tablenotebook"><td><p class = "paper">
+
 
−
June 3
+
<!---------------next day ----------------------------------------->
 +
 
 +
<tr class = "tablenotebook"><td><div class = "paper">
 +
&#9642; June 3
 
<br><br>
 
<br><br>
−
<strong>Yi Han</strong>
+
<span class = "brown"> Anaerobic Promoter</span>
 +
&emsp;&emsp;
 +
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Yi Han &#9728;</font>
 
<br><br>
 
<br><br>
−
&bull; Miniprep of gfp plasmids  
+
<div class = "divnbcontent">
−
<br>
+
<span class = "nbcontent">
−
&bull; RE digest with EcoRI and RsaI
+
Miniprep of gfp plasmids
−
</p>
+
</span><br>
 +
<span class = "nbcontent">
 +
RE digest with EcoRI and RsaI
 +
</span>
 +
</div>
 +
 
 +
<br><br>
 +
 
 +
</div>
 +
<div style = "border-top: 2px solid turquoise;"></div>
 
</td>
 
</td>
 
</tr>
 
</tr>
  
−
<tr class = "tablenotebook"><td><p class = "paper">
+
<!---------------next day ----------------------------------------->
−
June 4
+
 
 +
 
 +
<tr class = "tablenotebook"><td><div class = "paper">
 +
&#9642; June 4
 
<br><br>
 
<br><br>
−
<strong>Chi Yan</strong>
+
<span class = "brown"> Anaerobic Promoter</span>
 +
&emsp;&emsp;
 +
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Chi Yan &#9728;</font>
 
<br><br>
 
<br><br>
−
&bull; RE digest indicated a very faint smaller band for colonies 2-4, and hence these were likely to be positive clones
+
<div class = "divnbcontent">
−
<br>
+
<span class = "nbcontent">
−
</p>
+
RE digest indicated a very faint smaller band for colonies 2-4, and hence these were likely to be positive clones
 +
</span>
 +
</div>
 +
 
 +
<br><br>
 +
 
 +
</div>
 +
<div style = "border-top: 2px solid turquoise;"></div>
 
</td>
 
</td>
 
</tr>
 
</tr>
  
−
<tr class = "tablenotebook"><td><p class = "paper">
+
<!---------------next day ----------------------------------------->
−
June 5
+
 
 +
<tr class = "tablenotebook"><td><div class = "paper">
 +
&#9642; June 5
 
<br><br>
 
<br><br>
−
<strong>Yan Ting</strong>
+
<span class = "blue"> Invasin + Listerolysin</span>
 +
&emsp;&emsp;<span class = "black"> esa Quorum Sensing</span>
 +
<br>
 +
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Yan Ting &#9728;</font>
 
<br><br>
 
<br><br>
−
&bull; Inoculated 3mLS of Inv-4 and EsaR-4 plasmid carrying bacteria into 100mL LB+ appropriate Antibiotic
+
<div class = "divnbcontent">
 +
<span class = "nbcontent">
 +
Inoculated 3mLS of Inv-4 and EsaR-4 plasmid carrying bacteria into 100mL LB+ appropriate Antibiotic
 +
</span>
 
<br>
 
<br>
−
&bull; Cell culture &rarr; HEK293 cells revived from freezing down appeared detached.
+
<span class = "nbcontent">
−
<br>
+
Cell culture-> HEK293 cells revived from freezing down appeared detached.
−
</p>
+
</span>
 +
</div>
 +
 
 +
<br><br>
 +
 
 +
</div>
 +
<div style = "border-top: 2px solid turquoise;"></div>
 
</td>
 
</td>
 
</tr>
 
</tr>
  
−
<tr class = "tablenotebook"><td><p class = "paper">
+
 
−
June 6
+
<!---------------next day ----------------------------------------->
 +
 
 +
<tr class = "tablenotebook"><td><div class = "paper">
 +
&#9642; June 6
 
<br><br>
 
<br><br>
−
<strong>Yi Han & Yan Ting</strong>
+
<span class = "blue"> Invasin + Listerolysin</span>
 +
&emsp;&emsp;<span class = "black"> esa Quorum Sensing</span>
 +
&emsp;&emsp;<span class = "brown"> Anaerobic Promoter</span>
 +
<br>
 +
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Yi Han &#9728;</font>
 
<br><br>
 
<br><br>
−
&bull; Storage of bacterial glycerol stocks for Inv-4, EsaR4 in 25% glycerol
+
<div class = "divnbcontent">
 +
<span class = "nbcontent">
 +
Storage of bacterial glycerol stocks for Inv-4, EsaR4 in 25% glycerol
 +
</span>
 
<br>
 
<br>
−
&bull; Yan Ting: grew HEK293T cells in T25 glask
+
<span class = "nbcontent">
 +
Yanting: grew HEK293T cells in T25 flask
 +
</span>
 
<br>
 
<br>
−
&bull; Gel extract to clean up gfp plasmids which loading dye had accidentally been added to.
+
<span class = "nbcontent">
−
<br><br><br>
+
Gel extract to clean up gfp plasmids which loading dye had accidentally been added to.
−
</p>
+
</span>
 +
</div>
 +
 
 +
<br><br>
 +
 
 +
</div>
 +
<div style = "border-top: 2px solid turquoise;"></div>
 
</td>
 
</td>
 
</tr>
 
</tr>
 +
 +
<!---------------next day ----------------------------------------->
 +
 +
  
 
</table>
 
</table>
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<! ------------------------------- Footer START ------------------------------------------------>
 +
  
 
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<div class="left_sec">
 
<p>
 
<p>
−
<img src = "https://static.igem.org/mediawiki/2015/a/ab/SPSingapore_Team-Logo-New.png" height = "80px">
+
<img src = "https://static.igem.org/mediawiki/2015/a/ab/SPSingapore_Team-Logo-New.png" height = "60px" style="margin-top:5px;margin-left:-130px;">
 
</p>
 
</p>
 
</div>
 
</div>
 
<div class="right_sec">
 
<div class="right_sec">
 
<p>
 
<p>
−
<img src = "https://static.igem.org/mediawiki/2015/0/05/SPSingapore_NUS-Logo.png" height = "60px" style = "margin-top:10px;margin-right:30px;padding-right:30px;border-right: 3px solid lightgrey;">
+
<img src="https://static.igem.org/mediawiki/2015/0/05/SPSingapore_NUS-Logo.png" height="50px" style="margin-top:10px;margin-right:30px;padding-right:30px;border-right: 3px solid lightgrey;">
−
<img src = "https://static.igem.org/mediawiki/2015/7/72/SPSingapore_SPS-Logo.png" height = "60px" style = "margin-top:10px;margin-right:50px;"></a>
+
<img src="https://static.igem.org/mediawiki/2015/7/72/SPSingapore_SPS-Logo.png" height="50px" style="margin-top:10px;margin-right:30px;">
 
</p>
 
</p>
 
</div>
 
</div>

Latest revision as of 23:17, 18 September 2015


Research Notebook

Week 16 (6/9 - 12/9)

▪ May 31

Invasin + Listerolysin    esa Quorum Sensing
☀ Xin Yi ☀

Sent EsaR clone 4 and INv-4 for sequencing.


Invasin + Listerolysin    ☀ Yan Ting ☀

RE with XbaI and PstI
     RE - 2ul
     Buffer - 5ul
     DNA (4x of miniprep=74.5ng/ul) - 13.4ul
     H2O - 29.6ul
     Incubate at 37degC for 2.5h (1130-1400)

▶ Results: Gel loaded 1kb ladder, uncut, RE digested. 100V for 1h.
Plasmid doesn't have XbaI site - RE digested DNA is a linear 6kb band.


▪ June 2

Anaerobic Promoter    ☀ Xin Yi ☀

YFP and GFP transformation results - no colonies for YFP
The GFP transformation repeated with 100ng of plasmid was sucessful. 4 colonies of gfp plasmid were inoculated in 3mL LB+amp and grown overnight.


▪ June 3

Anaerobic Promoter    ☀ Yi Han ☀

Miniprep of gfp plasmids
RE digest with EcoRI and RsaI


▪ June 4

Anaerobic Promoter    ☀ Chi Yan ☀

RE digest indicated a very faint smaller band for colonies 2-4, and hence these were likely to be positive clones


▪ June 5

Invasin + Listerolysin    esa Quorum Sensing
☀ Yan Ting ☀

Inoculated 3mLS of Inv-4 and EsaR-4 plasmid carrying bacteria into 100mL LB+ appropriate Antibiotic
Cell culture-> HEK293 cells revived from freezing down appeared detached.


▪ June 6

Invasin + Listerolysin    esa Quorum Sensing    Anaerobic Promoter
☀ Yi Han ☀

Storage of bacterial glycerol stocks for Inv-4, EsaR4 in 25% glycerol
Yanting: grew HEK293T cells in T25 flask
Gel extract to clean up gfp plasmids which loading dye had accidentally been added to.