From 2013.igem.org

(Difference between revisions)

Revision as of 01:56, 28 September 2013

导航栏

Biobrick Transformation (6.24-6.30)

2013/9/26 20:16:17--Chun Tang

2013/9/26 20:16:39

2013/9/26 20:17:26

Plasmids Construction (7.1-7.28, 8.15-9.16)

2013/9/26 20:36:39--Chun Tang

2013/9/26 20:36:58

2013/9/28 01:03:53

2013/9/26 20:37:51--Chun Tang

2013/9/26 20:38:17

2013/9/28 01:05:10

2013/9/28 01:05:15

2013/9/26 20:40:10--Chun Tang

2013/9/26 20:40:27

2013/9/28 01:06:41

2013/9/28 01:06:51

2013/9/26 20:43:04--Chun Tang

2013/9/26 20:43:20

2013/9/28 01:10:10

2013/9/28 01:10:38

2013/9/26 20:44:56--Chun Tang

2013/9/26 20:45:14

2013/9/28 01:11:54

2013/9/28 01:12:42

2013/9/26 20:47:01--Chun Tang

2013/9/26 20:47:15

2013/9/28 01:16:48

2013/9/28 01:17:27

2013/9/26 20:48:37--Chun Tang

2013/9/26 20:48:50

2013/9/28 01:19:02

Transformation (7.29-9.20)

2013/9/26 20:49:56--Chun Tang

2013/9/26 20:50:04

2013/9/28 01:21:56

2013/9/28 01:22:37

2013/9/26 20:51:34--Chun Tang

2013/9/26 20:51:47

2013/9/28 01:23:53

2013/9/26 20:52:47--Chun Tang

2013/9/26 20:53:01

2013/9/28 01:24:59

2013/9/26 20:55:54--Chun Tang

2013/9/26 20:56:19

SDS-PAGE

2013/9/26 21:48:57--Tang Chun

2013/9/26 21:49:44

2013/9/26 21:50:08--Tang Chun

2013/9/26 21:51:07

2013/9/26 21:53:17--Tang Chun

2013/9/26 21:55:45

2013/9/26 22:02:08--Tang Chun

2013/9/26 22:03:34

2013/9/26 22:04:18--Tang Chun

2013/9/26 22:07:07

2013/9/26 22:07:33--Tang Chun

2013/9/26 22:08:04

2013/9/26 22:07:46

2013/9/26 22:08:31--Tang Chun

2013/9/26 22:08:47

2013/9/26 22:09:10--Tang Chun

2013/9/26 22:09:28

2013/9/26 22:09:40

2013/9/26 22:10:13--Tang Chun

2013/9/26 22:10:54

2013/9/26 22:11:28

2013/9/26 22:12:08--Tang Chun

2013/9/26 22:12:23

2013/9/26 22:12:32

2013/9/26 22:13:05--Tang Chun

2013/9/26 22:13:16

2013/9/28 07:03:20--Chun Tang

2013/9/28 07:03:54

2013/9/28 08:44:02--Chun Tang

2013/9/28 08:44:22

2013/9/28 08:47:11

2013/9/28 08:44:24

2013/9/28 08:44:28

Microfluidic Experiment Record

2013/9/18 10:50:50--Chun Tang

2013/9/25 20:38:47

2013/9/18 10:51:01--Chun Tang

2013/9/25 20:42:38

2013/9/18 10:51:41--Chun Tang

2013/9/25 20:45:15

2013/9/18 10:52:00--Chun Tang

2013/9/25 20:46:04

2013/9/18 10:52:31--Chun Tang

2013/9/25 20:47:42

2013/9/18 10:53:02--Chun Tang

2013/9/25 20:50:35

2013/9/18 10:53:24--Chun Tang

2013/9/25 20:52:02

2013/9/25 21:10:34--Chun Tang

2013/9/25 21:11:06

2013/9/25 21:12:48--Chun Tang

2013/9/25 21:13:01

2013/9/28 07:14:25

2013/9/28 07:14:27

2013/9/28 07:14:28

2013/9/25 21:15:00--Chun Tang

2013/9/25 21:15:47

2013/9/25 21:16:23

2013/9/25 21:16:51

RFU (9.24-9.27)

2013/9/27 21:59:07--Chun Tang

2013/9/27 21:59:17

2013/9/28 03:11:44

2013/9/28 03:30:57

2013/9/28 03:31:38

2013/9/28 03:34:06

@@ Line 167: / Line 167: @@
 <h1>SDS-PAGE</h1>
 				<div class="M1_exp_stepInfo" title="Abbreviation:" id="sb7s0" style="background-color: rgb(166, 166, 166);">			<button class="M1_exp_step_name_button" style="background-color: rgb(166, 166, 166);"><input value="Abbreviation:" class="M1_exp_step_name"></button>			<span class="M1_exp_step_time">2013/9/26 21:48:57--Tang Chun</span>		</div>		<table class="M1_exp_stepBoard">		<tbody><tr class="M1_exp_stepItem">			<td class="M1_exp_item_time" id="ib7s0i0">2013/9/26 21:49:44</td>			<td class="M1_exp_item_content"><textarea id="i7s0i0" class="C0" rows="1" disabled="" style="background-color: white;">W: Whole cell   P: Precipitation   S: Supernatant
-</textarea></td>		</tr>		</tbody></table>				<div class="M1_exp_stepInfo" title="Background" id="sb7s1" style="background-color: rgb(166, 166, 166);">			<button class="M1_exp_step_name_button" style="background-color: rgb(166, 166, 166);"><input value="Background" class="M1_exp_step_name"></button>			<span class="M1_exp_step_time">2013/9/26 21:50:08--Tang Chun</span>		</div>		<table class="M1_exp_stepBoard">		<tbody><tr class="M1_exp_stepItem">			<td class="M1_exp_item_time" id="ib7s1i0">2013/9/26 21:51:07</td>			<td class="M1_exp_item_content"><table id="i7s1i0" class="C3"><tbody><tr><td class="C3_td"><input value="CDS" class="C3_input"></td><td class="C3_td"><input value="Size(bp)" class="C3_input"></td><td class="C3_td"><input value="Protein" class="C3_input"></td><td class="C3_td"><input value="Size(kDa)" class="C3_input"></td></tr><tr><td class="C3_td"><input value="luxR" class="C3_input"></td><td class="C3_td"><input value="750" class="C3_input"></td><td class="C3_td"><input value="LuxR" class="C3_input"></td><td class="C3_td"><input value="27.5" class="C3_input"></td></tr><tr><td class="C3_td"><input value="gfp-lva" class="C3_input"></td><td class="C3_td"><input value="753" class="C3_input"></td><td class="C3_td"><input value="GFP-LVA" class="C3_input"></td><td class="C3_td"><input value="27.61" class="C3_input"></td></tr><tr><td class="C3_td"><input value="aiiA-lva" class="C3_input"></td><td class="C3_td"><input value="783" class="C3_input"></td><td class="C3_td"><input value="aiiA-LVA" class="C3_input"></td><td class="C3_td"><input value="28.71" class="C3_input"></td></tr><tr><td class="C3_td"><input value="luxI-lva" class="C3_input"></td><td class="C3_td"><input value="612" class="C3_input"></td><td class="C3_td"><input value="LuxI-LVA" class="C3_input"></td><td class="C3_td"><input value="22.44" class="C3_input"></td></tr><tr><td class="C3_td"><input value="ndh-lva" class="C3_input"></td><td class="C3_td"><input value="1335" class="C3_input"></td><td class="C3_td"><input value="NDH-LVA" class="C3_input"></td><td class="C3_td"><input value="48.95" class="C3_input"></td></tr></tbody></table></td>		</tr>		</tbody></table>				<div class="M1_exp_stepInfo" title="Graphic Symbol" id="sb7s2" style="background-color: rgb(166, 166, 166);">			<button class="M1_exp_step_name_button" style="background-color: rgb(166, 166, 166);"><input value="Graphic Symbol" class="M1_exp_step_name"></button>			<span class="M1_exp_step_time">2013/9/26 21:53:17--Tang Chun</span>		</div>		<table class="M1_exp_stepBoard">		<tbody><tr class="M1_exp_stepItem">			<td class="M1_exp_item_time" id="ib7s2i0">2013/9/26 21:55:45</td>			<td class="M1_exp_item_content"><table id="i7s2i0" class="C3"><tbody><tr><td class="C3_td"><input value="Strain" class="C3_input"></td><td class="C3_td"><input value="Plasmids" class="C3_input"></td><td class="C3_td"><input value="Proteins" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21" class="C3_input"></td><td class="C3_td"><input value="/" class="C3_input"></td><td class="C3_td"><input value="NDH" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-A" class="C3_input"></td><td class="C3_td"><input value="pSB1C3-gfp-luxI" class="C3_input"></td><td class="C3_td"><input value="Luxr,LuxI-LVA,GFP-LVA" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-B" class="C3_input"></td><td class="C3_td"><input value="pSB3T5-aiiA" class="C3_input"></td><td class="C3_td"><input value="/" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-C" class="C3_input"></td><td class="C3_td"><input value="pSB4K5-ndh" class="C3_input"></td><td class="C3_td"><input value="/" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-A B" class="C3_input"></td><td class="C3_td"><input value="pSB1C3-gfp-luxI pSB3T5-aiiA" class="C3_input"></td><td class="C3_td"><input value="Luxr,LuxI-LVA,GFP,aiiA-LVA" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-A B C" class="C3_input"></td><td class="C3_td"><input value="pSB1C3-gfp luxI pSB3T5-aiiA pSB4K5-ndh" class="C3_input"></td><td class="C3_td"><input value="LuxR,LuxI-LVA,GFP-LVA.aiiA-LVA.NDH-LVA" class="C3_input"></td></tr><tr><td class="C3_td"><input value="" class="C3_input"></td><td class="C3_td"><input value="" class="C3_input"></td><td class="C3_td"><input value="" class="C3_input"></td></tr></tbody></table></td>		</tr>		</tbody></table>				<div class="M1_exp_stepInfo" title="1. Characterization of expression of three single plasmids in BL21. (8.02-8. 08)" id="sb7s3" style="background-color: rgb(166, 166, 166);">			<button class="M1_exp_step_name_button" style="background-color: rgb(166, 166, 166);"><input value="1. Characterization of expression of three single plasmids in BL21. (8.02-8. 08)" class="M1_exp_step_name"></button>			<span class="M1_exp_step_time">2013/9/26 22:02:08--Tang Chun</span>		</div>		<table class="M1_exp_stepBoard">		<tbody><tr class="M1_exp_stepItem">			<td class="M1_exp_item_time" id="ib7s3i0">2013/9/26 22:03:34</td>			<td class="M1_exp_item_content">	<table id="i7s3i0" class="C1">		<tbody><tr>			<td class="C1_td">				<img id="i7s3i00" src="http://trysomething-block.stor.sinaapp.com/t14i9s3i01.png"><br>				<form id="i7s3i04" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form>				<input name="img" type="file" id="i7s3i02" form="i7s3i04" class="C1_button1" required="required" disabled="" style="background-color: white;">				<input name="pname" type="hidden" value="t14" form="i7s3i04">				<input name="id" type="hidden" value="i7s3i0" form="i7s3i04">				<input name="oldgraph" type="hidden" id="i7s3i05" form="i7s3i04" value="t14i9s3i01.png">				<input id="i7s3i06" type="submit" value="submit" form="i7s3i04" class="C1_button2" disabled="" style="background-color: white;">				<div id="i7s3i03" class="M1_exp_img"></div>				</td>			<td class="C1_td_text">				<textarea id="i7s3i01" class="C1_text" disabled="" style="background-color: white;">  We compared protein bands between BL21, the blank control and three groups transformed into different single plasmids, aiiA, ndh and gfp. These two gel pictures were got in two different days.
+</textarea></td>		</tr>		</tbody></table>				<div class="M1_exp_stepInfo" title="Background" id="sb7s1" style="background-color: rgb(166, 166, 166);">			<button class="M1_exp_step_name_button" style="background-color: rgb(166, 166, 166);"><input value="Background" class="M1_exp_step_name"></button>			<span class="M1_exp_step_time">2013/9/26 21:50:08--Tang Chun</span>		</div>		<table class="M1_exp_stepBoard">		<tbody><tr class="M1_exp_stepItem">			<td class="M1_exp_item_time" id="ib7s1i0">2013/9/26 21:51:07</td>			<td class="M1_exp_item_content"><table id="i7s1i0" class="C3"><tbody><tr><td class="C3_td"><input value="CDS" class="C3_input"></td><td class="C3_td"><input value="Size(bp)" class="C3_input"></td><td class="C3_td"><input value="Protein" class="C3_input"></td><td class="C3_td"><input value="Size(kDa)" class="C3_input"></td></tr><tr><td class="C3_td"><input value="luxR" class="C3_input"></td><td class="C3_td"><input value="750" class="C3_input"></td><td class="C3_td"><input value="LuxR" class="C3_input"></td><td class="C3_td"><input value="27.5" class="C3_input"></td></tr><tr><td class="C3_td"><input value="gfp-lva" class="C3_input"></td><td class="C3_td"><input value="753" class="C3_input"></td><td class="C3_td"><input value="GFP-LVA" class="C3_input"></td><td class="C3_td"><input value="27.61" class="C3_input"></td></tr><tr><td class="C3_td"><input value="aiiA-lva" class="C3_input"></td><td class="C3_td"><input value="783" class="C3_input"></td><td class="C3_td"><input value="aiiA-LVA" class="C3_input"></td><td class="C3_td"><input value="28.71" class="C3_input"></td></tr><tr><td class="C3_td"><input value="luxI-lva" class="C3_input"></td><td class="C3_td"><input value="612" class="C3_input"></td><td class="C3_td"><input value="LuxI-LVA" class="C3_input"></td><td class="C3_td"><input value="22.44" class="C3_input"></td></tr><tr><td class="C3_td"><input value="ndh-lva" class="C3_input"></td><td class="C3_td"><input value="1335" class="C3_input"></td><td class="C3_td"><input value="NDH-LVA" class="C3_input"></td><td class="C3_td"><input value="48.95" class="C3_input"></td></tr></tbody></table></td>		</tr>		</tbody></table>				<div class="M1_exp_stepInfo" title="Graphic Symbol" id="sb7s2" style="background-color: rgb(166, 166, 166);">			<button class="M1_exp_step_name_button" style="background-color: rgb(166, 166, 166);"><input value="Graphic Symbol" class="M1_exp_step_name"></button>			<span class="M1_exp_step_time">2013/9/26 21:53:17--Tang Chun</span>		</div>		<table class="M1_exp_stepBoard">		<tbody><tr class="M1_exp_stepItem">			<td class="M1_exp_item_time" id="ib7s2i0">2013/9/26 21:55:45</td>			<td class="M1_exp_item_content"><table id="i7s2i0" class="C3"><tbody><tr><td class="C3_td"><input value="Strain" class="C3_input"></td><td class="C3_td"><input value="Plasmids" class="C3_input"></td><td class="C3_td"><input value="Proteins" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21" class="C3_input"></td><td class="C3_td"><input value="/" class="C3_input"></td><td class="C3_td"><input value="NDH" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-A" class="C3_input"></td><td class="C3_td"><input value="pSB1C3-gfp-luxI" class="C3_input"></td><td class="C3_td"><input value="Luxr,LuxI-LVA,GFP-LVA" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-B" class="C3_input"></td><td class="C3_td"><input value="pSB3T5-aiiA" class="C3_input"></td><td class="C3_td"><input value="/" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-C" class="C3_input"></td><td class="C3_td"><input value="pSB4K5-ndh" class="C3_input"></td><td class="C3_td"><input value="/" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-A B" class="C3_input"></td><td class="C3_td"><input value="pSB1C3-gfp-luxI pSB3T5-aiiA" class="C3_input"></td><td class="C3_td"><input value="Luxr,LuxI-LVA,GFP,aiiA-LVA" class="C3_input"></td></tr><tr><td class="C3_td"><input value="BL21-A B C" class="C3_input"></td><td class="C3_td"><input value="pSB1C3-gfp luxI pSB3T5-aiiA pSB4K5-ndh" class="C3_input"></td><td class="C3_td"><input value="LuxR,LuxI-LVA,GFP-LVA.aiiA-LVA.NDH-LVA" class="C3_input"></td></tr><tr><td class="C3_td"><input value="" class="C3_input"></td><td class="C3_td"><input value="" class="C3_input"></td><td class="C3_td"><input value="" class="C3_input"></td></tr></tbody></table></td>		</tr>		</tbody></table>				<div class="M1_exp_stepInfo" title="1. Characterization of expression of three single plasmids in BL21. (8.02-8. 08)" id="sb7s3" style="background-color: rgb(166, 166, 166);">			<button class="M1_exp_step_name_button" style="background-color: rgb(166, 166, 166);"><input value="1. Characterization of expression of three single plasmids in BL21. (8.02-8. 08)" class="M1_exp_step_name"></button>			<span class="M1_exp_step_time">2013/9/26 22:02:08--Tang Chun</span>		</div>		<table class="M1_exp_stepBoard">		<tbody><tr class="M1_exp_stepItem">			<td class="M1_exp_item_time" id="ib7s3i0">2013/9/26 22:03:34</td>			<td class="M1_exp_item_content">	<table id="i7s3i0" class="C1">		<tbody><tr>			<td class="C1_td">				<img id="i7s3i00" width="500px" src="http://trysomething-block.stor.sinaapp.com/t14i9s3i01.png"><br>				<form id="i7s3i04" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form>				<input name="img" type="file" id="i7s3i02" form="i7s3i04" class="C1_button1" required="required" disabled="" style="background-color: white;">				<input name="pname" type="hidden" value="t14" form="i7s3i04">				<input name="id" type="hidden" value="i7s3i0" form="i7s3i04">				<input name="oldgraph" type="hidden" id="i7s3i05" form="i7s3i04" value="t14i9s3i01.png">				<input id="i7s3i06" type="submit" value="submit" form="i7s3i04" class="C1_button2" disabled="" style="background-color: white;">				<div id="i7s3i03" class="M1_exp_img"></div>				</td>			<td class="C1_td_text">				<textarea id="i7s3i01" class="C1_text" disabled="" style="background-color: white;">  We compared protein bands between BL21, the blank control and three groups transformed into different single plasmids, aiiA, ndh and gfp. These two gel pictures were got in two different days.
 Data analysis:
-   The target proteins of pSB3T5-aiiA/BL21 and pSB4K5-ndh/BL21 should not be expressed because the promoter should be induced by AHL. The target proteins of pSB1C3-gfp-luxI/BL21 have been expressed but GFP was misfolded we suppose. However, the band marked in red was not the target band. Maybe it's the complex of AHL-Lux</textarea>				<br>			</td>		</tr>	</tbody></table>	</td>		</tr>		</tbody></table>				<div class="M1_exp_stepInfo" title="2. Characterization of expression of double plasmids (8.05)" id="sb7s4" style="background-color: rgb(166, 166, 166);">			<button class="M1_exp_step_name_button" style="background-color: rgb(166, 166, 166);"><input value="2. Characterization of expression of double plasmids (8.05)" class="M1_exp_step_name"></button>			<span class="M1_exp_step_time">2013/9/26 22:04:18--Tang Chun</span>		</div>		<table class="M1_exp_stepBoard">		<tbody><tr class="M1_exp_stepItem">			<td class="M1_exp_item_time" id="ib7s4i0">2013/9/26 22:07:07</td>			<td class="M1_exp_item_content">	<table id="i7s4i0" class="C1">		<tbody><tr>			<td class="C1_td">				<img id="i7s4i00" src="http://trysomething-block.stor.sinaapp.com/t14i9s4i02.png"><br>				<form id="i7s4i04" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form>				<input name="img" type="file" id="i7s4i02" form="i7s4i04" class="C1_button1" required="required" disabled="" style="background-color: white;">				<input name="pname" type="hidden" value="t14" form="i7s4i04">				<input name="id" type="hidden" value="i7s4i0" form="i7s4i04">				<input name="oldgraph" type="hidden" id="i7s4i05" form="i7s4i04" value="t14i9s4i02.png">				<input id="i7s4i06" type="submit" value="submit" form="i7s4i04" class="C1_button2" disabled="" style="background-color: white;">				<div id="i7s4i03" class="M1_exp_img"></div>				</td>			<td class="C1_td_text">				<textarea id="i7s4i01" class="C1_text" disabled="" style="background-color: white;">  We performed a SDS-PAGE comparing expression of double plasmids, gfp and aiiA, and two corresponding single plasmids. What's more, we wanted to find more details about band which possibly is the misfolded gfp.
+   The target proteins of pSB3T5-aiiA/BL21 and pSB4K5-ndh/BL21 should not be expressed because the promoter should be induced by AHL. The target proteins of pSB1C3-gfp-luxI/BL21 have been expressed but GFP was misfolded we suppose. However, the band marked in red was not the target band. Maybe it's the complex of AHL-LuxI.</textarea>				<br>			</td>		</tr>	</tbody></table>	</td>		</tr>		</tbody></table>				<div class="M1_exp_stepInfo" title="2. Characterization of expression of double plasmids (8.05)" id="sb7s4" style="background-color: rgb(166, 166, 166);">			<button class="M1_exp_step_name_button" style="background-color: rgb(166, 166, 166);"><input value="2. Characterization of expression of double plasmids (8.05)" class="M1_exp_step_name"></button>			<span class="M1_exp_step_time">2013/9/26 22:04:18--Tang Chun</span>		</div>		<table class="M1_exp_stepBoard">		<tbody><tr class="M1_exp_stepItem">			<td class="M1_exp_item_time" id="ib7s4i0">2013/9/26 22:07:07</td>			<td class="M1_exp_item_content">	<table id="i7s4i0" class="C1">		<tbody><tr>			<td class="C1_td">				<img id="i7s4i00" src="http://trysomething-block.stor.sinaapp.com/t14i9s4i02.png"><br>				<form id="i7s4i04" target="M7" action="img.php" method="post" enctype="multipart/form-data"></form>				<input name="img" type="file" id="i7s4i02" form="i7s4i04" class="C1_button1" required="required" disabled="" style="background-color: white;">				<input name="pname" type="hidden" value="t14" form="i7s4i04">				<input name="id" type="hidden" value="i7s4i0" form="i7s4i04">				<input name="oldgraph" type="hidden" id="i7s4i05" form="i7s4i04" value="t14i9s4i02.png">				<input id="i7s4i06" type="submit" value="submit" form="i7s4i04" class="C1_button2" disabled="" style="background-color: white;">				<div id="i7s4i03" class="M1_exp_img"></div>				</td>			<td class="C1_td_text">				<textarea id="i7s4i01" class="C1_text" disabled="" style="background-color: white;">  We performed a SDS-PAGE comparing expression of double plasmids, gfp and aiiA, and two corresponding single plasmids. What's more, we wanted to find more details about band which possibly is the misfolded gfp.
 Data analysis:
    The band marked in red disappeared is pSB1C3-gfp-luxI/BL21.The LuxR, GFP and AiiA  are the same of size (marked in yellow). However the band of AiiA was not clear. Many target proteins were misfolded. Therefore the culture condition should be optimized.

Team:XMU-China/wetlab journal