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Team:Hong Kong HKUST/wetlab/notebook
From 2014.igem.org
(Difference between revisions)
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| - | <div id="satu" align="center"> <h2><center>July | + | <div id="satu" align="center"> <h2><center>July 2014</center></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> July | + | <br> <b> July 2</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| - | · <br> | + | · Arrival of primers order for comX-myc and comW-FLAG<br> |
| + | · Dilution of primers and design of experiment for both comX-myc and comW-FLAG<br> | ||
| + | · Inoculation of ligated pSB1C3-K880005.B0015<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 3</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · PCR of comX-myc using Taq DNA Polymerase with ThermoPol Buffer to check whether primers are functional<br> | ||
| + | · Gel check for comX-myc PCR product (PCR product present)<br> | ||
| + | · PCR of comX-myc using Phusion High-Fidelity PCR Master Mix with HF Buffer<br> | ||
| + | · Gel check for comX-myc PCR product (no PCR product present)<br> | ||
| + | · Restriction digestion check on some candidate colonies of ligated pSB1C3-K880005.B0015 with XbaI and PstI<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 4</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · PCR of comX-myc using Vent DNA Polymerase, PCR clean up<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 5</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | <br> <b> July 6</b> <br> | ||
| + | Wet lab<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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<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> July | + | <br> <b> July 7</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Digestion of comX-myc with XbaI and PstI<br> | ||
| + | · Gel check of digested comX-myc, digestion clean up<br> | ||
| + | · PCR of comW-FLAG using Vent DNA Polymerase<br> | ||
| + | · Gel check for comW-FLAG PCR product, PCR clean up<br> | ||
| + | · Dephosporylation of digested pSB1C3-K880005 backbone cut with SpeI and PstI<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 8</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Ligation of comX-myc cut with XbaI and PstI into pSB1C3-K880005 backbone cut with SpeI and PstI<br> | ||
| + | · 2nd attempt of ligation of BBa_B0015 insert cut with XbaI and PstI into dephosporylated pSB1C3-K880005 backbone cut with SpeI and PstI as negative control<br> | ||
| + | · Transformation of ligation products<br> | ||
| + | · <br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 9</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Inoculation of ligated pSB1C3-K880005.B0015 (2nd attempt) and ligated pSB1C3-K880005.comX-myc<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 10</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Another PCR of comW-FLAG using Vent DNA Polymerase<br> | ||
| + | · Gel check for comW-FLAG PCR product, PCR clean up<br> | ||
| + | · Restriction digestion check on some candidate colonies of ligated pSB1C3-K880005.B0015 (2nd attempt) and ligated pSB1C3-K880005.comX-myc<br> | ||
| + | · Gel check and insert wanted present in several candidate colonies of ligated pSB1C3-K880005.B0015 (2nd attempt), but no insert wanted present in any candidate colonies of ligated pSB1C3-K880005.comX-myc<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | <br> <b> July 11</b> <br> | ||
| + | Wet lab<br> | ||
| + | · Digestion of comW-FLAG with XbaI and PstI, digestion clean up<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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<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> July | + | <br> <b> July 14</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Ligation of comW-FLAG cut on XbaI and PstI into dephosporylated pSB1C3-K880005 backbone cut on SpeI and PstI<br> | ||
| + | · Transformation of ligation products<br> | ||
| + | · Inoculation of ligated dephosporylated pSB1C3-K880005.comX-myc<br> | ||
| + | · <br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 15</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| - | · <br> | + | · Digestion check of ligated pSB1C3-K880005.comX-myc<br> |
| + | · Digestion of pSB1C3-K880005.comX-myc on SpeI and PstI, but no insert wanted present<br> | ||
| + | · PCR of comX using Vent DNA Polymerase, PCR clean-up<br> | ||
| + | · Colony PCR of pSB1C3-K880005.comX-myc using Taq DNA Polymerase<br> | ||
| + | · Gel check of pSB1C3-K880005.comX-myc PCR product (no PCR product present)<br> | ||
| + | · Inoculation of dephosporylated pSB1C3-K880005.comX-myc, dephosporylated pSB1C3-K880005.comW-FLAG<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 16</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Digestion check of ligated pSB1C3-K880005.comW-FLAG<br> | ||
| + | · Colony PCR of pSB1C3-K880005.comW-FLAG<br> | ||
| + | · Gel check of pSB1C3-K880005.comW-FLAG PCR product (no PCR product present)<br> | ||
| + | · PCR of comW using Vent DNA Polymerase (no PCR product present)<br> | ||
| + | · Digestion of comX on XbaI and PstI<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 17</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · PCR of comX-myc and comX using Vent DNA Polymerase<br> | ||
| + | · Gel check of comX-myc and comX PCR product, PCR clean-up<br> | ||
| + | · PCR of comW using Vent DNA Polymerase<br> | ||
| + | · Gel extraction and gel purification of comW PCR product<br> | ||
| + | · Dephosporylation of BBa_K880005<br> | ||
· <br> | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | <br> <b> July 18</b> <br> | ||
| + | Wet lab<br> | ||
| + | · PCR of comX using Vent DNA Polymerase<br> | ||
| + | · Gel check of comX PCR product, PCR clean-up<br> | ||
| + | · Digestion of comX PCR product on XbaI and PstI, digestion clean-up<br> | ||
| + | · Digestion of comX-myc and comW PCR products, digestion clean-up<br> | ||
| + | · Gel extraction and gel purification of digested comX and comX-myc<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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<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> July | + | <br> <b> July 21</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| - | · <br> | + | · Gel purification of comX and comX-myc<br> |
| + | · Ligation of comX-myc into dephosporylated pSB1C3-K880005 backbone with 1:4, 1:7, 1:10 ratio<br> | ||
| + | · Ligation of comW-FLAG into dephosporylated pSB1C3-K880005 backbone with 1:4, 1:7, 1:10 ratio<br> | ||
| + | · Ligation of comX into dephosporylated pSB1C3-K880005 backbone with 1:3 and 1:5 ratio<br> | ||
| + | · Ligation of comW into dephosporylated pSB1C3-K880005 backbone with 1:4, 1:7, 1:10 ratio<br> | ||
| + | · Colony PCR of pSB1C3-K880005.comW-FLAG using Taq DNA Polymerase<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 22</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Colony PCR of comX, comW, comX-myc, comW-FLAG and use BBa_J04450 as the positive control using Taq DNA Polymerase (no PCR product present)<br> | ||
| + | · Inoculation of comX, comW, comX-myc, comW-FLAG<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 23</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Digestion check of comX, comW, comX-myc, comW-FLAG colony PCR products on XbaI and SpeI<br> | ||
| + | · Inoculation of comW-FLAG<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 24</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Digestion check of pSB1C3-K880005.comW-FLAG on XbaI and SpeI-HF<br> | ||
| + | · Ligation of comW-FLAG into dephosporylated pSB1C3-K880005 backbone<br> | ||
| + | · Ligation of comX-myc into dephosporylated pSB1C3-K880005 backbone<br> | ||
| + | · PCR of comX-myc and comX using Vent DNA Polymerase, gel extraction and gel purification<br> | ||
| + | · Digestion of comX-myc and comX on XbaI and PstI-HF<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | <br> <b> July 25</b> <br> | ||
| + | Wet lab<br> | ||
| + | · Colony PCR of comX-myc using Taq DNA Polymerase<br> | ||
| + | · Gel check of comX-myc PCR product (no PCR product present)<br> | ||
| + | · <br> | ||
| + | · <br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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<a href="#" class="head">Week 5</a> | <a href="#" class="head">Week 5</a> | ||
<div class="content" align="left" style="display: none;"> | <div class="content" align="left" style="display: none;"> | ||
| - | <br> <b> July | + | <br> <b> July 28</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Colony PCR of comX-myc using Taq DNA Polymerase<br> | ||
| + | · Colony PCR of comw-FLAG using Taq DNA Polymerase<br> | ||
| + | · Inoculation of comX-myc and comW-FLAG<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 29</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Digestion check of comX-myc and comW-FLAG on XbaI and PstI-HF<br> | ||
| + | · Transformation of comX-myc and comW-FLAG<br> | ||
| + | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 30</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| + | · Digestion check of comX-myc and comW-FLAG<br> | ||
| + | · Inoculation of comX-myc and comW-FLAG <br> | ||
| + | · Streaking plates of comX-myc and comW-FLAG<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
| - | <br> <b> July | + | <br> <b> July 31</b> <br> |
Wet lab<br> | Wet lab<br> | ||
| - | · <br> | + | · Digestion check of comX-myc and comW-FLAG on XbaI and PstI-HF<br> |
| + | · Redigest comW-FLAG on XmnI and NcoI<br> | ||
| + | · PCR sequencing for comW and comX<br> | ||
| + | · Ethanol precipitation for comW and comX<br> | ||
· <br> | · <br> | ||
· <br> | · <br> | ||
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| - | <div id="satu" align="center"> <h2>August | + | <div id="satu" align="center"> <h2>August 2014</h2> |
<div> | <div> | ||
Revision as of 15:29, 6 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. PneumosensorRiboregulator |
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