Team:Hong Kong HKUST/wetlab/notebook

From 2014.igem.org

(Difference between revisions)
Line 603: Line 603:
-
<div id="satu" align="center"> <h2>August 2013</h2>
+
<div id="satu" align="center"> <h2>August 2014</h2>
    
    
<div>
<div>
       <a href="#" class="head">Week 1</a>
       <a href="#" class="head">Week 1</a>
       <div class="content" align="left" style="display: none;">
       <div class="content" align="left" style="display: none;">
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 1</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Sequencing of comX-myc and comW-FLAG<br>
 +
·      Digestion of BBa_B0015 plasmid on EcoRI-HF and XbaI<br>
 +
·      Digestion pSB1C3-K880005.comX-myc on EcoRI-HF and SpeI, digestion clean-up<br>
 +
·      Digestion pSB1C3-K880005.comX-myc on PstI-HF and SpeI, digestion clean-up<br>
·      <br>
·      <br>
 +
·      <br>
 +
·   <br>
 +
<br><b>Aug 4</b> <br>
 +
Wet lab<br>
 +
·      Digestion of comX-myc on EcoRI-HF and SpeI, gel extraction and gel purification<br>
 +
·      Ligation of BBa_B0015 cut on XbaI and PstI into pSB1C3-comX-myc backbone cut on SpeI and PstI<br>
 +
·      Transformation of pSB1C3-comX-myc.B0015 ligation product<br>
 +
·      Ligation of comX-myc cut on EcoRI-HF and SpeI into pSB1C3-B0015 backbone cut on EcoRI-HF and XbaI<br>
 +
·      Transformation of pSB1C3-B0015.comX-myc ligation product<br>
 +
·      Sequencing of comX-myc and comW-FLAG<br>
 +
·      Inoculation of comX-myc<br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 5</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Sequencing result failed<br>
 +
·      Transformation of pSB1C3-B0015.comX-myc ligation product failed<br>
 +
·      Digestion check of comX-myc on StyI<br>
 +
·      Colony PCR of comX-myc<br>
 +
·      Inoculation of comX-myc<br>
·      <br>
·      <br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
 +
·   <br>
 +
<br><b>Aug 6</b> <br>
 +
Wet lab<br>
 +
·      Digestion check of comX-myc on XbaI and PstI<br>
 +
·      Gel check of comX-myc PCR product (no PCR product present)<br>
 +
·      Sequencing of comW-FLAG using VF2 and VR<br>
 +
·      Digestion of comX-myc on StyI<br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 7</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion of comX-myc on StyI<br>
 +
·      Digestion check of comX on SphI and NcoI<br>
 +
·      Digestion check of comW on XmnI and NcoI<br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 8</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion of comW-FLAG on EcoRI-HF and SpeI, gel extraction and gel purification<br>
 +
·      Digestion of comW-FLAG on SpeI and PstI-HF, digestion clean-up<br>
 +
·      Sequencing of comX-myc succeeded but mutation at EcoRI site<br>
 +
·      Digestion check of comX-myc at EcoRI-HF and PstI-HF<br>
 +
·      Ligation of comX and comW into dephosporylated pSB1C3-K880005<br>
 +
·      Transformation of comX and comW ligation product<br>
 +
·      Digestion of comX-myc at EcoRI-HF and SpeI-HF<br>
 +
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
Line 639: Line 680:
       <a href="#" class="head">Week 2</a>
       <a href="#" class="head">Week 2</a>
       <div class="content" align="left" style="display: none;">
       <div class="content" align="left" style="display: none;">
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 11</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion check of comX-myc and comW-FLAG on XbaI and PstI-HF<br>
 +
·      Gel purification of comX-myc digested on EcoRI-HF and SpeI<br>
 +
·      Ligation of comX-myc cut on EcoRI-HF and SpeI into pSB1C3-B0015 cut on EcoRI-HF and XbaI<br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 12</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion check of pSB1C3-K880005.comX and pSB1C3-K880005.comW-FLAG on XbaI and PstI-HF<br>
 +
·      Dilution of comW-FLAG<br>
 +
·      Re-digestion of BBa_B0015 on EcoRI-HF and XbaI, digestion clean-up<br>
 +
·      Re-ligation of pSB1C3-B0015.comX-myc<br>
 +
·      Transformation of pSB1C3-B0015.comX-myc ligation product<br>
 +
·      Inoculation of pSB1C3-K880005.comX-myc.B0015 and pSB1C3-K880005.comX-myc<br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 13</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion check of pSB1C3-B0015.comX-myc and pSB1C3-K880005.comX-myc.B0015 on XbaI and PstI-HF<br>
 +
·      <br>
 +
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 14</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion check of pSB1C3-K880005.comX-myc.B0015 on XbaI and PstI-HF<br>
 +
·      Transformation of ligated product pSB1C3-K880005.comX-myc.B0015<br>
 +
·      Dilution of comW-FLAG<br>
 +
·      Digestion of comW-FLAG on EcoRI-HF and SpeI-HF<br>
 +
·      Ligation of comW-FLAG into pSB1C3-B0015 backbone<br>
 +
·      Re-digestion of comW-FLAG on EcoRI-HF and SpeI-HF<br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
 +
·   <br>
 +
<br><b>Aug 15</b> <br>
 +
Wet lab<br>
 +
·      Digestion of pSB1C3-K880005 on SpeI-HF and PstI-HF, digestion clean-up<br>
 +
·      Colony PCR of comW-FLAG using Taq DNA Polymerase<br>
 +
·      Digestion of pSB1C3-B0015 on EcoRI-HF and XbaI<br>
 +
·      Digestion of pSB1C3-K880005.comX-myc on EcoRI-HF and SpeI-HF<br>
 +
·      Digestion of pSB1C3-B0015 on EcoRI-HF and XbaI, digestion clean-up<br>
 +
·      Digestion of pSB1C3-K880005.comX-myc on EcoRI-HF and SpeI-HF, gel extraction and gel purification<br>
 +
·      Overnight ligation of BBa_B0015 into pSB1C3-K880005.comX-myc backbone at 16°C<br>
 +
·      Transformation of pSB1C3-K880005.comX-myc.B0015 ligation product<br>
 +
·      Ligation of comX-myc into pSB1C3-B0015 backbone<br>
 +
·      Inoculation of pSB1C3-K880005.comW-FLAG and pSB1C3-K880005.comX-myc.B0015<br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
 
       </div>
       </div>
Line 670: Line 747:
       <a href="#" class="head">Week 3</a>
       <a href="#" class="head">Week 3</a>
       <div class="content" align="left" style="display: none;">
       <div class="content" align="left" style="display: none;">
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 18</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Transformation of ligated pSB1C3-K880005.comX-myc<br>
 +
·      Ligation of comW-FLAG into pSB1C3-K880005 backbone, comX-myc into pSB1C3-K880005 backbone, BBa_B0015 into pSB1C3-K880005.comX-myc backbone<br>
 +
·      Digestion check of pSB1C3-K880005.comW-FLAG on EcoRI-HF and SpeI-HF<br>
 +
·      Digestion check of pSB1C3-K880005.comW-FLAG on SpeI-HF and PstI-HF<br>
 +
·      Digestion of pSB1C3-K880005.comX-myc on EcoRI-HF and SpeI-HF, gel extraction and gel purification<br>
 +
·      Inoculation of pSB1C3-K880005.comX-myc and pSB1C3-B0015<br>
 +
·      Digestion of pSB1C3-K880005.comX-myc on SpeI-HF and PstI-HF, digestion clean-up<br>
 +
·      Digestion check of pSB1C3-K880005.comX-myc.B0015 on XbaI and PstI-HF<br>
 +
·      Digestion of pSB1C3-K880005 on SpeI-HF and PstI-HF, digestion clean-up<br>
 +
·      <br>
 +
·      <br>
 +
·   <br>
 +
<br><b>Aug 19</b> <br>
 +
Wet lab<br>
 +
·      Digestion of pSB1C3-B0015 on EcoRI-HF and XbaI, digestion clean-up<br>
 +
·      Digestion check of comX-myc on XbaI and PstI-HF<br>
 +
·      Ligation of pSB1C3-K880005.comX-myc cut on EcoRI-HF and SpeI-HF iinto pSB1C3-B0015 backbone cut on EcoRI-HF and XbaI<br>
 +
·      Inoculation of pSB1C3-K880005.comW-FLAG and pSB1C3-K880005.comX-myc.B0015<br>
 +
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 20</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion check of pSB1C3-K880005.comW-FLAG on EcoRI-HF and SpeI-HF, gel extraction and gel purification<br>
 +
·      Digestion check of pSB1C3-K880005.comX-myc.B0015 on XbaI and PstI-HF<br>
 +
·      Digestion check of pSB1C3-K880005.comX-myc.B0015 on SphI and SpeI-HF<br>
 +
·      Digestion check of pSB1C3-K880005.comX-myc on XbaI and PstI<br>
 +
·      Inoculation of pSB1C3-K880005.comX-myc.B0015 and pSB1C3-K880005.comW-FLAG<br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 21</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion check of pSB1C3-K880005.comW-FLAG on EcoRI-HF and SpeI-HF<br>
 +
·      Digestion check of pSB1C3-K880005.comX-myc.B0015 on XbaI and PstI-HF<br>
 +
·      Ligation of pSB1C3-K880005.comW-FLAG cut on EcoRI-HF and SpeI-HF into pSB1C3-B0015 backbone cut on EcoRI-HF and XbaI<br>
 +
·      Colony PCR of pSB1C3-B0015 and reverse comX-myc using Taq DNA Polymerase<br>
 +
·      Digestion of comW-FLAG on NotI-HF<br>
 +
·      Digestion of comW-FLAG on EcoRI-HF and SpeI-HF<br>
 +
·      Inoculation of pSB1C3-K880005.comW-FLAG<br>
 +
·      Transformation pSB1C3-K880005.comW-FLAG ligation product and pSB1C3-K880005.comX-myc.B0015<br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 22</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion check of comW-FLAG on EcoRI-HF and SpeI-HF<br>
 +
·      Digestion of pSB1C3-K880005.comX-myc.B0015 on EcoRI-HF and SpeI-HF<br>
 +
·      Inoculation pSB1C3-K880005.comX-myc.B0015<br>
 +
·      <br>
 +
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
Line 702: Line 819:
       <a href="#" class="head">Week 4</a>
       <a href="#" class="head">Week 4</a>
       <div class="content" align="left" style="display: none;">
       <div class="content" align="left" style="display: none;">
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 25</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Ligation of pSB1C3-K880005.comW-FLAG cut on EcoRI-HF and SpeI-HF into pSB1C3-B0015 backbone cut on EcoRI-HF and XbaI<br>
 +
·      Digestion check of pSB1C3-K880005.comX-myc.B0015 on XbaI and PstI-HF<br>
 +
·      Transformation of pSB1C3-K880005.comW-FLAG<br>
 +
·      Digestion of pSB1C3-K880005.comX-myc.B0015 on SpeI-HF and PstI-HF, gel extraction and gel purification<br>
 +
·      Inoculation of pSB1C3-K880005.comX-myc.B0015 for glycerol stock<br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 26</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion of pSB1C3-K880005.comX-myc.B0015 on SpeI-HF and PstI-HF, digestion clean-up<br>
 +
·      Ligation of Combox + GFP cut on XbaI and PstI-HF into pSB1C3-K880005.comX-myc.B0015 backbone cut on SpeI-HF and PstI-HF<br>
 +
·      Digestion of BBa_E0240 100 bp on XbaI and PstI-HF, gel extraction and gel purification<br>
 +
·      Inoculation of pSB1C3-K880005.comW-FLAG.B0015<br>
 +
·      Transformation of pSB1C3-K880005.comW-FLAG<br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 27</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion check of pSB1C3-K880005.comW-FLAG on XbaI and PstI-HF, gel extraction and gel purification<br>
 +
·      Sequencing of pSB1C3-K880005.comW-FLAG.B0015<br>
 +
·      Ligation of pSB1C3-K880005.comW-FLAG into pSB1C3-B0015 backbone and Combox + GFP into pSB1C3-K880005.comX-myc.B0015 backbone<br>
 +
·      Transformation of pSB1C3-K880005.comW-FLAG<br>
 +
·      Inoculation of pSB1C3-K880005.comW-FLAG.B0015 and pSB1C3-K880005.comW-FLAG<br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>
·   <br>
·   <br>
-
<br><b>Aug xx</b> <br>
+
<br><b>Aug 28</b> <br>
Wet lab<br>
Wet lab<br>
 +
·      Digestion check of pSB1C3-K880005.comW-FLAG.B0015 on XbaI and PstI-HF, gel extraction and gel purification<br>
 +
·      Digestion check of pSB1C3-K880005.comW-FLAG on EcoRI-HF and SpeI-HF, gel extraction and gel purification<br>
 +
·      Ligation of Combox + GFP into pSB1C3-K880005.comX-myc.B0015 backbone<br>
 +
·      Inoculation of pSB1C3-K880005.comW-FLAG.B0015 and pSB1C3-K880005.comW-FLAG<br>
 +
·      Ligation of pSB1C3-K880005.comW-FLAG into pSB1C3-B0015 backbone<br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
 +
·   <br>
 +
<br><b>Aug 29</b> <br>
 +
Wet lab<br>
 +
·      Digestion check of pSB1C3-K880005.comW-FLAG.B0015 on XbaI and PstI-HF<br>
 +
·      Digestion check of pSB1C3-K880005.comW-FLAG on EcoRI-HF and SpeI-HF<br>
 +
·      <br>
 +
·      <br>
 +
·      <br>
·      <br>
·      <br>
·      <br>
·      <br>

Revision as of 14:29, 3 October 2014


Notebook

Throughout the course of our project, we have continuously recorded our daily work, such as wetlab activities and research findings. The notebook for Pneuumonsensor and Riboregulator are displayed separately.






Pneumosensor

June 2014

Week 1
Week 2
Week 3
Week 4
Week 5

July 2014

Week 1
Week 2
Week 3
Week 4
Week 5

August 2014

Week 1
Week 2
Week 3
Week 4
Week 5

September 2013

Week 1
Week 2
Week 3
Week 4
Week 5





Riboregulator

June 2013

Week 1
Week 2
Week 3
Week 4
Week 5

July 2013

Week 1
Week 2
Week 3
Week 4
Week 5

August 2013

Week 1
Week 2
Week 3
Week 4
Week 5

September 2013

Week 1
Week 2
Week 3
Week 4
Week 5

Home

Team

Main Project

Side Project

Human Practice

Wetlab

Achievement