Template:Kyoto/Notebook/Sep 2

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Revision as of 06:40, 23 September 2013

Sep 2

Liquid Culture

Hirano

Sample	medium
9/1 entA-(Master Plate)-1	4mL SOB(+Km)

37°C

Colony PCR

Tatsui

Sample	base pair
9/1 RBS-lysis2-DT-(1)	985
Ptet-RBS-lacZα-DT-(1)	765
Plac-RBS-lacZα-DT-(1)	765
Plac-RBS-lacZα-DT-(2)	765

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5min	30s	30s	1min	30cycles

File:Igku sep2electrophoresis1.png

Restriction Enzyme Digestion

Nakamoto

	pSB1C3-(1)	EcoRI	SpeI	XbaI	PstI	BufferB	BufferD	BSA	MilliQ	total
2 cuts(E+S)	7µL	1µL	1µL	0µL	0µL	3µL	0µL	0.3µL	17.7µL	30µL
NC(E+S)	0.3µL	0µL	0µL	0µL	0µL	1µL	0µL	0.1µL	8.6µL	10µL
2 cut(X+P)s	7µL	0µL	0µL	1µL	1µL	0µL	3µL	0.3µL	17.7µL	30µL
NC(X+P)	0.3µL	0µL	0µL	0µL	0µL	0µL	1µL	0.1µL	8.6µL	10µL

	8/21 tRMA-spinach(1)	EcoRI	SpeI	BufferB	BSA	MilliQ	total
2 cuts	8µL	1.0µL	1.0µL	3µL	0.3µL	17.6µL	30µL
NC	0.5µL	0µL	0µL	1µL	0.1µL	8.4µL	10µL

	8/21 tetR aptamer12_1R-(1)	EcoRI	SpeI	BufferB	BSA	MilliQ	total
2 cuts	6µL	1.0µL	1.0µL	3µL	0.3µL	18.7µL	30µL
NC	0.4µL	0µL	0µL	1µL	0.1µL	8.5µL	10µL

	8/21 pT181 attenuator-(2)	EcoRI	SpeI	XbaI	PstI	BufferB	BufferD	BSA	MilliQ	total
2 cuts(E+S)	8µL	1µL	1µL	0µL	0µL	3µL	0µL	0.3µL	16.3µL	30µL
NC(E+S)	0.5µL	0µL	0µL	0µL	0µL	1µL	0µL	0.1µL	8.4µL	10µL
2 cut(X+P)s	8µL	0µL	0µL	1µL	1µL	0µL	3µL	0.3µL	16.3µL	30µL
NC(X+P)	0.5µL	0µL	0µL	0µL	0µL	0µL	1µL	0.1µL	8.4µL	10µL

Liquid Culture

Kojima

Sample	medium
tRNA-Spinach-1	Plusgrow medium(+CP)
tetR aptamaer12_1R-1	Plusgrow medium(+CP)
pT181 attenuator-1	Plusgrow medium(+CP)	=	pT181 antisense-1	Plusgrow medium(+CP)

Master Plate

Kojima

Number	Use LB plate(+CP)
1	tRNA Spinach-1
2	tetR aptamaer12_1R
3	tetR aptamaer12_P
4	tetR aptamaer12_1M
5	pT181 attenuator-2
6	Fusion1 attenuator-1
7	Fusion3m2 attenuator-1
8	pT181 antisense-1
9	Fusion1 antisense-1
10	Fuaion6 antisense-1

Gel Extraction

No name

Lane	DNA	Enzyme
1	100bp ladder	-
2	pSB1C3(E+S)	EcoRI&SpeI
3	pSB1C3(E+S)	EcoRI&SpeI
4	--	--
5	8/24 tetR aptamer 12_1R	EcoRI&SpeI
6	8/24 tetR aptamer 12_1R	EcoRI&SpeI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
No name

Lane	DNA	Enzyme
1	100bp ladder	-
2	pT181 attenuator(2)(E+S)	EcoRI&SpeI
3	pT181 attenuator(2)(E+S)	EcoRI&SpeI
4	--	--
5	pT181 attenuator(2)(X+P)	EcoRI&SpeI
6	pT181 attenuator(2)(X+P)	EcoRI&SpeI
7	--	--
8	pT181 attenuator(2)(X+P)	EcoRI&SpeI
9	pT181 attenuator(2)(X+P)	EcoRI&SpeI
10	--	--
11	Spinach	EcoRI&SpaI
12	Spinach	EcoRI&SpaI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

Name	concentration[µg/mL]	260/280	260/230
pSB1C3 (EcoRI&SpeI)	3.0	2.18	0.31
pSB1C3 (XbaI&PstI)	4.7	2.25	0.36
pT181 attenuator-(2)(EcoR&SpeI)	8.6	2.74	0.01
pT181 attenuator-(2) (XbaI&PstI)	16.5	2.46	0.03
Spinach (EcoRI&SpeI)	2.8	2.98	0.27
tetR aptamer12_1R (EcoRI&SpeI)	50.4	28.07	1.97

Colony PCR

No name

Sample	base pair
8/27 pT181 attenuator-DT-1	738
8/27 pT181 attenuator-DT-2	738
8/27 pT181 attenuator-DT-3	738
8/27 TetR-aptamer 12_1R-DT-1	521
8/27 TetR-aptamer 12_1R-DT-2	521
8/27 pT181 antisense-DT-1	542

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5min	30s	30s	30s	30cycles

File:Igku Aug28electrophoresis3?.png

Sample	base pair
8/27 Pλ-luxI-1	???
8/27 Pλ-luxI	???

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5min	30s	30s	30min	3cycles

File:Igku Aug19electrophoresis1.png

Master Plate

Hirano

Liquid Culture

Hirano

Colony PCR

Hirano

Transformation

Hirano

@@ Line 187: / Line 187: @@
 |}
 </div>
-===EDTA Solution===
-<div class="experiment">
-<span class="author">Hirano</span>
 ===Colony PCR===
 <div class="experiment">
-<span class="author">Hirano</span>
+<span class="author">No name</span>
+{| class="wikitable"
+!|Sample||base pair
+|-
+|8/27 pT181 attenuator-DT-1||738
+|-
+|8/27 pT181 attenuator-DT-2||738
+|-
+|8/27 pT181 attenuator-DT-3||738
+|-
+|8/27 TetR-aptamer 12_1R-DT-1||521
+|-
+|8/27 TetR-aptamer 12_1R-DT-2||521
+|-
+|8/27 pT181 antisense-DT-1||542
+|}
+{| class="wikitable"
+!PreDenature||Denature||Annealing||Extension||cycle
+|-
+|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
+|-
+|5min||30s||30s||30s||30cycles
+|}
+[[File:igku_Aug28electrophoresis3?.png]]
+{| class="wikitable"
+!|Sample||base pair
+|-
+|8/27 P&lambda;-luxI-1||???
+|-
+|8/27 P&lambda;-luxI||???
+|}
+{| class="wikitable"
+!PreDenature||Denature||Annealing||Extension||cycle
+|-
+|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
+|-
+|5min||30s||30s||30min||3cycles
+|}
+[[File:igku_Aug19electrophoresis1.png]]
+</div>
 ===Master Plate===

Template:Kyoto/Notebook/Sep 2

From 2013.igem.org

Revision as of 06:40, 23 September 2013

Contents

Sep 2

Liquid Culture

Colony PCR

Restriction Enzyme Digestion

Liquid Culture

Master Plate

Gel Extraction

Colony PCR

Master Plate

Liquid Culture

Colony PCR

Transformation