Biomod/2012/TeamSendai/Experiment - Revision history

Kazuki Hirahara at 04:00, 28 October 2012

Kazuki Hirahara — Sun, 28 Oct 2012 04:00:44 GMT

←Older revision		Revision as of 04:00, 28 October 2012
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	[[Image:スクリーンショット 2012-10-28 11.09.52.png\|800px\|thumb\|left\|SPR analysis]]		[[Image:スクリーンショット 2012-10-28 11.09.52.png\|800px\|thumb\|left\|SPR analysis]]
-	[[Image:スクリーンショット 2012-10-28 12.55.51.png\|800px\|thumb\|left\|SPR analysis Zoom]]	+	[[Image:スクリーンショット 2012-10-28 12.59.02.png\|800px\|thumb\|left\|SPR analysis Zoom]]

Kazuki Hirahara at 03:58, 28 October 2012

Kazuki Hirahara — Sun, 28 Oct 2012 03:58:38 GMT

←Older revision		Revision as of 03:58, 28 October 2012
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	<h3>Results</h3>		<h3>Results</h3>
-	From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we could insert min-gate into liposome by attaching cholesterol-leg.

-	From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels (> 100?差を記述！) than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we could insert the gate into lipid bilayer membrane by attaching cholesterol-leg.

-	~~[[Image:スクリーンショット 2012~~-10-~~28 11~~.09.52.~~png\|800px\|thumb\|left\|SPR analysis]]~~	+	From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels 22.4 than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we could insert the gate into lipid bilayer membrane by attaching cholesterol-leg.

		+	[[Image:スクリーンショット 2012-10-28 11.09.52.png\|800px\|thumb\|left\|SPR analysis]]
		+	[[Image:スクリーンショット 2012-10-28 12.55.51.png\|800px\|thumb\|left\|SPR analysis Zoom]]

Kazuki Hirahara at 03:56, 28 October 2012

Kazuki Hirahara — Sun, 28 Oct 2012 03:56:24 GMT

←Older revision		Revision as of 03:56, 28 October 2012
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	Inject sample 10 μL and start measurement. After end of measurement, inject 10 mM NaOH 5 μL. It is necessary to unify measurement time and RU levels when inject sample.		Inject sample 10 μL and start measurement. After end of measurement, inject 10 mM NaOH 5 μL. It is necessary to unify measurement time and RU levels when inject sample.

-	Sample 1:	+	Sample 1　consists　on　that:
-		+
	min-gate (4.2 nM) 50 μL		min-gate (4.2 nM) 50 μL

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	Running buffer 38 μL		Running buffer 38 μL

-	~~Sample2~~:	+	Sample　2　consists on that:

	min-gate 50 (4.2 nM) μL		min-gate 50 (4.2 nM) μL
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	<h3>Results</h3>		<h3>Results</h3>
	From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we could insert min-gate into liposome by attaching cholesterol-leg.		From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we could insert min-gate into liposome by attaching cholesterol-leg.
		+
		+	From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels (> 100?差を記述！) than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we could insert the gate into lipid bilayer membrane by attaching cholesterol-leg.

	[[Image:スクリーンショット 2012-10-28 11.09.52.png\|800px\|thumb\|left\|SPR analysis]]		[[Image:スクリーンショット 2012-10-28 11.09.52.png\|800px\|thumb\|left\|SPR analysis]]

Naito Ushio at 03:24, 28 October 2012

Naito Ushio — Sun, 28 Oct 2012 03:24:06 GMT

←Older revision		Revision as of 03:24, 28 October 2012
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	From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we could insert min-gate into liposome by attaching cholesterol-leg.		From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we could insert min-gate into liposome by attaching cholesterol-leg.

-	[[Image:スクリーンショット 2012-10-28 11.09.52.png\|~~550px~~\|thumb\|left\|SPR analysis]]	+	[[Image:スクリーンショット 2012-10-28 11.09.52.png\|800px\|thumb\|left\|SPR analysis]]
-		+
-		+
-		+
-		+
-		+
-		+
-		+


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		+	{{-}}

	<h3>References</h3>		<h3>References</h3>

Naito Ushio at 03:21, 28 October 2012

Naito Ushio — Sun, 28 Oct 2012 03:21:24 GMT

←Older revision		Revision as of 03:21, 28 October 2012
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	Next,we tried to create the GATE which can be decorated with cholesterol to connect it with liposome. We call this tube “connect-able GATE”.		Next,we tried to create the GATE which can be decorated with cholesterol to connect it with liposome. We call this tube “connect-able GATE”.
-	[[Image:imashun8.png\|thumb\|center\|400px\|GATE ~~decolated~~ with cholesterol]]	+	[[Image:imashun8.png\|thumb\|center\|400px\|GATE decorated with cholesterol]]

	When we annealed the connect-able GATE and did electrophoresis, the band could be seen at the similar position compared with normal GATE. For this, we consider the connect-able GATE also may be created and we could find many granular objects when we observed the connect-able GATE by AFM.		When we annealed the connect-able GATE and did electrophoresis, the band could be seen at the similar position compared with normal GATE. For this, we consider the connect-able GATE also may be created and we could find many granular objects when we observed the connect-able GATE by AFM.

Naito Ushio at 03:18, 28 October 2012

Naito Ushio — Sun, 28 Oct 2012 03:18:37 GMT

←Older revision		Revision as of 03:18, 28 October 2012
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	Next,we tried to create the GATE which can be decorated with cholesterol to connect it with liposome. We call this tube “connect-able GATE”.		Next,we tried to create the GATE which can be decorated with cholesterol to connect it with liposome. We call this tube “connect-able GATE”.
-	[[Image:imashun8.png\|thumb\|center\|400px]]	+	[[Image:imashun8.png\|thumb\|center\|400px\|GATE decolated with cholesterol]]

	When we annealed the connect-able GATE and did electrophoresis, the band could be seen at the similar position compared with normal GATE. For this, we consider the connect-able GATE also may be created and we could find many granular objects when we observed the connect-able GATE by AFM.		When we annealed the connect-able GATE and did electrophoresis, the band could be seen at the similar position compared with normal GATE. For this, we consider the connect-able GATE also may be created and we could find many granular objects when we observed the connect-able GATE by AFM.
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	<h3>Results</h3>		<h3>Results</h3>
-	From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we ~~can~~ insert min-gate into liposome by attaching cholesterol-leg.	+	From the graph, we confirmed that min-gate with cholesterol-leg had higher RU levels than only min-gate. It shows that min-gate with cholesterol-leg has higher strength of binding. Therefore, we could insert min-gate into liposome by attaching cholesterol-leg.

	[[Image:スクリーンショット 2012-10-28 11.09.52.png\|550px\|thumb\|left\|SPR analysis]]		[[Image:スクリーンショット 2012-10-28 11.09.52.png\|550px\|thumb\|left\|SPR analysis]]

Naito Ushio at 03:14, 28 October 2012

Naito Ushio — Sun, 28 Oct 2012 03:14:04 GMT

←Older revision		Revision as of 03:14, 28 October 2012
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	Chloroform 70μL		Chloroform 70μL

-	We ~~dry~~ a sample of that composition under argon gas condition.	+	We dried a sample of that composition under argon gas condition.

	For liposome formation, 125μL of 1×TAE　Mg<sup>2+</sup> were added and incubate for 1 h at room temperature.		For liposome formation, 125μL of 1×TAE　Mg<sup>2+</sup> were added and incubate for 1 h at room temperature.

Naito Ushio at 03:12, 28 October 2012

Naito Ushio — Sun, 28 Oct 2012 03:12:48 GMT

←Older revision		Revision as of 03:12, 28 October 2012
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	[[Image:Porterbeforesensyoku.png\|thumb\|left\|300px\|fig.1 Lanes are 1: target and Porter1, 2: target and Porter1 and Porter2, 3: target and Porter1 and Porter2 and Porter3]][[Image:Poteraftersensyoku.png\|thumb\|none\|360px\|fig.2 stained samples]]		[[Image:Porterbeforesensyoku.png\|thumb\|left\|300px\|fig.1 Lanes are 1: target and Porter1, 2: target and Porter1 and Porter2, 3: target and Porter1 and Porter2 and Porter3]][[Image:Poteraftersensyoku.png\|thumb\|none\|360px\|fig.2 stained samples]]

-		+	{{-}}
	If the target has passed well through Porters, bands of the target hybridizing with Porter1, 2, and 3 should appear. In the gel, bands above each Porter can be seen as the Porter hybridizing with the target. <br>		If the target has passed well through Porters, bands of the target hybridizing with Porter1, 2, and 3 should appear. In the gel, bands above each Porter can be seen as the Porter hybridizing with the target. <br>
	In the first 15 minutes, Porter1 attached to the target (fig.1: lane 3). After another 15 minutes, the band of Porter1 and Porter2 hybridizing with the target were seen. In the last 15 minutes, the band of Porter1 and Porter2 and Porter3 were seen.<br>		In the first 15 minutes, Porter1 attached to the target (fig.1: lane 3). After another 15 minutes, the band of Porter1 and Porter2 hybridizing with the target were seen. In the last 15 minutes, the band of Porter1 and Porter2 and Porter3 were seen.<br>
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	<h2>Confirming insertion of the Gate s into the membrane</h2>		<h2>Confirming insertion of the Gate s into the membrane</h2>

-	We ~~use~~ fluoresce moelcules to confirm that the tube insert into the liposome. If the mini-gate were inserted into membrane and make a hole on membrane, inner solution without negative charges should be leaked. Lucifer Yellow fluorescein were encapsulated into giant liposome. Leaking was measured by comparing fluorescent intensity between inside and outside of liposome.As a positive control of the leaking assay, alpha- Hemolysin protein was used. alpha- Hemolysin protein is known to make a hole in liposomes.	+	We used fluoresce moelcules to confirm that the tube insert into the liposome. If the mini-gate were inserted into membrane and make a hole on membrane, inner solution without negative charges should be leaked. Lucifer Yellow fluorescein were encapsulated into giant liposome. Leaking was measured by comparing fluorescent intensity between inside and outside of liposome.As a positive control of the leaking assay, alpha- Hemolysin protein was used. alpha- Hemolysin protein is known to make a hole in liposomes.
	The following figure summarizes the leaking assay.		The following figure summarizes the leaking assay.

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	Yellow line area is a domain of a graph expressed by ''"FigureB2'"'		Yellow line area is a domain of a graph expressed by ''"FigureB2'"'
	]]		]]
-	[[Image:スクリーンショット 2012-10-27 23.21.03.png\|350px\|thumb\|center\|''FigureB2'' Gray scale graph]]	+	[[Image:スクリーンショット 2012-10-27 23.21.03.png\|350px\|thumb\|center\|''FigureB2'' Gray scale graph
	----		----
	This is the gray scale graph using ''ImageJ'' soft.The area of		This is the gray scale graph using ''ImageJ'' soft.The area of

Naito Ushio at 03:07, 28 October 2012

Naito Ushio — Sun, 28 Oct 2012 03:07:44 GMT

←Older revision		Revision as of 03:07, 28 October 2012
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	<h4>Results</h4>		<h4>Results</h4>
-	[[Image:~~Picture1-1-1~~.png\|thumb\|center\|650px\|alt=this is Electrophpresis\|Fig 3. Lanes are 1: Porter1, 2: Porter2, 3: Target, 4: toehold A, 5: toehold B, 6: target and Porter1, 7: target and Porter1 and 2, 8: target and Porter2, 9: target and toehold A, 10: target and toehold A and toehold B, 11: target and toehold B, 12: 20b ladder	+	[[Image:Poterwik線入りi.png\|thumb\|center\|650px\|alt=this is Electrophpresis\|Fig 3. Lanes are 1: Porter1, 2: Porter2, 3: Target, 4: toehold A, 5: toehold B, 6: target and Porter1, 7: target and Porter1 and 2, 8: target and Porter2, 9: target and toehold A, 10: target and toehold A and toehold B, 11: target and toehold B, 12: 20b ladder
	The target was passed between porters (lane 7). But the target didn’t attach to toehold A (lane 9). So, the target cannot be delivered by the toehold structure (lane 10)]]		The target was passed between porters (lane 7). But the target didn’t attach to toehold A (lane 9). So, the target cannot be delivered by the toehold structure (lane 10)]]

Kazuki Hirahara at 02:43, 28 October 2012

Kazuki Hirahara — Sun, 28 Oct 2012 02:43:56 GMT

←Older revision		Revision as of 02:43, 28 October 2012
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	{{:Biomod/2012/Tohoku/Team_Sendai/header}}		{{:Biomod/2012/Tohoku/Team_Sendai/header}}