Template:Kyoto/Notebook/Aug 30

From 2013.igem.org

(Difference between revisions)

Revision as of 12:15, 22 September 2013

Aug 30

Gel Extraction

No Name

Name	quantity	concentration[µg/mL]	260/280	260/230
pT181 attenuator(2) (EcoRI & SpeI)	305.3	2.2	1.66	0.36
pT181 attenuator(2) (XbaI & PstI)	217.7	3.9	1.73	0.04
pT181 antisense(2) (EcoRI & SpeI)	347.7	3.1	1.88	0.31
pT181 antisense(2) (XbaI & PstI)	376.5	5.9	0.49	2.36
pSB1C3 (EcoRI & SpeI)	436.6	30.7	1.03	1.28
pSB1C3 (XbaI & PstI)	457.4	7.9	1.65	0.42
Spinach (EcoRI & SpeI)	490.5	3.0	1.87	0.26
tetR aptamer 12_1R (EcoRI & SpeI)	451.7	-12.8	5.56	-0.04

Miniprep

No name

DNA	concentration[µg/mL]	260/280	260/230
RBS-lysis2 9	264.6	1.97	2.21
RBS-lysis2 12	220.0	1.89	2.46
RBS-lysis2 14	195.3	1.88	1.53

Colony PCR

No name

Sample	base pair
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT(8/20 LB Amp) (1)~(4)	2143
8/29 Plux-RBS-GFP-DT(8/27 LB-CP) (1)~(2)	1227
8/29 Pbad/araC-RBS-RFP{BBa_I13516} (8/21 LB-CP) (1)~(2)	2256
8/29 RBS-lysis3-DT-(1)	1210

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5min	30s	30s	2min	30cycle

Ethanol Precipitation

No name

DNA
pT181 attenuator-(2) (EcoRI & SpeI)
pT181 attenuator-(2) (XbaI & PstI)
pT181 antisense-(2) (EcoRI & SpeI)
pT181 antisense-(2) (XbaI & PstI)
pSB1C3 (EcoRI & SpeI)
pSB1C3 (XbaI & PstI)
Spinach (EcoRI & SpeI)
tetR aptamer 12_1R (EcoRI & SpeI)

Restriction Enzyme Digestion

No name

	8/30 RBS-lysis2 (9)	EcoRI	SpeI	10x bufferB	100x BSA	MilliQ	total
2 cuts	7.5 µL	0.5 µL	0.5 µL	3 µL	0.3 µL	18.2 µL	30 µL
NC	0.5 µL	0 µL	0 µL	1 µL	0.1 µL	8.4 µL	10 µL

	8/22 pSB1C3-(2)	EcoRI	SpeI	10x Buffer	100x BSA	MilliQ	total
2 cuts	8 µL	0.5 µL	0.5 µL	3 µL	0.3 µL	17.7 µL	30 µL
NC	1 µL	0 µL	0 µL	1 µL	0.1 µL	7.9 µL	10 µL

	8/22 pSB1C3-(2)	XbaI	PstI	BufferD	BSA	MilliQ	total
2 cuts	8 µL	0.5 µL	0.5 µL	3 µL	0.3 µL	17.7 µL	30 µL
NC	1 µL	0 µL	0 µL	1 µL	0.1 µL	7.9 µL	10 µL

	8/21 Spinach (1)	EcoRI	SpeI	10x BufferB	100x BSA	MilliQ	total
2 cuts	4 µL	0.5 µL	0.5 µL	3 µL	0.3 µL	21.9 µL	30 µL
NC	0.5 µL	0 µL	0 µL	1 µL	0.1 µL	8.4 µL	10 µL

	8/21 tetR aptamer 12_1R (1)	EcoRI	SpeI	10xBufferB	100xBSA	MilliQ	total
2 cuts	3 µL	0.5 µL	0.5 µL	3 µL	0.3 µL	22.7 µL	30 µL
NC	0.4 µL	0 µL	0 µL	1 µL	0.1 µL	8.4 µL	10 µL

	8/21 pT181 attenuator-(2)	EcoRI	SpeI	XbaI	PstI	10x BufferB	10x BufferD	100x BSA	MilliQ	total
2 cuts	4 µL	0.5 µL	0.5 µL	0 µL	0 µL	3 µL	0 µL	0.3 µL	21.7 µL	30 µL
NC	0.5 µL	0 µL	0 µL	0 µL	0 µL	1 µL	0 µL	0.1 µL	8.4 µL	10 µL
2 cuts	4 µL	0 µL	0 µL	0.5 µL	0.5 µL	0 µL	3 µL	0.3 µL	21.7 µL	30 µL
NC	0.5 µL	0 µL	0 µL	0 µL	0 µL	0 µL	1 µL	0.1 µL	8.4 µL	10 µL

	8/21 pT181 antisense-(2)	EcoRI	SpeI	XbaI	PstI	10x BufferB	10x BufferD	100x BSA	MilliQ	total
2 cuts	9 µL	0.5 µL	0.5 µL	0 µL	0 µL	3 µL	0 µL	0.3 µL	16.7 µL	30 µL
NC	1 µL	0 µL	0 µL	0 µL	0 µL	1 µL	0 µL	0.1 µL	7.9 µL	10 µL
2 cuts	9 µL	0 µL	0 µL	0.5 µL	0.5 µL	0 µL	3 µL	0.3 µL	16.7 µL	30 µL
NC	1 µL	0 µL	0 µL	0 µL	0 µL	0 µL	1 µL	0.1 µL	7.9 µL	10 µL

Electrophoresis

No name

Lane	Sample	Enzyme1	Enzyme2
1	100bp ladder	--	--
2	RBS-lysis2-(9)	EcoRI	SpeI
3	RBS-lysis2-(9)	--	--
4	pSB1C3-(2)	EcoRI	SpeI
5	pSB1C3-(2)	--	--
6	pSB1C3-(2)	XbaI	PstI
7	pSB1C3-(2)	--	--
8	tRNA-Spinach-(1)	EcoRI	SpeI
9	tRNA-Spinach-(1)	--	--
10	tetR aptamer12_1R	EcoRI	SpeI
11	tetR aptamer12_1R	--	--
12	pT181 attenuator	EcoRI	SpeI
13	pT181 attenuator	EcoRI	SpeI
14	100bp ladder	--	--

File:Igku Aug30electrophoresis1

Lane	Sample	Enzyme1	Enzyme2
1	100bp ladder	--	--
2	pT181 antisense	EcoRI	SpeI
3	pT181 antisense	--	--
4	pT181 antisense	XbaI	PstI
5	pT181 antisense	--	--
6	pT181 attenuator	XbaI	PstI
7	pT181 attenuator	--	--
8	100bp ladder	--	--

File:Igku Aug30electrophoresis2

Lane	Sample
1	1kb ladder
2	8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)(Colony PCR product)
3	8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)(Colony PCR product)
4	8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)(Colony PCR product)
5	8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)(Colony PCR product)
6	Pbad/araC-RBS-RFP-DT-(1)(Colony PCR product)
7	Pbad-araC-RBS-RFP-DT-(2)(Colony PCR product)

File:Igku Aug30electrophoresis3

Liquid Culture

No name

Sample	medium
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)	Plusgrow medium(+Amp)
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)	Plusgrow medium(+Amp)
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)	Plusgrow medium(+Amp)
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)	Plusgrow medium(+Amp)
8/29 Plux-RBS-GFP-DT-(1)	Plusgrow medium(+CP)
8/29 Plux-RBS-GFP-DT-(2)	Plusgrow medium(+CP)
8/29 Pbad/araC-RBS-RFP-DT-(1)	Plusgrow medium(+CP)
8/29 Pbad-araC-RBS-RFP-DT-(2)	Plusgrow medium(+CP)
RBS-lysis3-DT-(1)	Plusgrow medium(+CP)

@@ Line 221: / Line 221: @@
 |}
 [[File:igku_Aug30electrophoresis3]]<br>
+===Liquid Culture===
+<!-- ここから -->
+<div class="experiment">
+<span class="author">No name</span>
+{| class="wikitable"
+!Sample||medium
+|-
+|8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)||Plusgrow medium(+Amp)
+|-
+|8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)||Plusgrow medium(+Amp)
+|-
+|8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)||Plusgrow medium(+Amp)
+|-
+|8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)||Plusgrow medium(+Amp)
+|-
+|8/29 Plux-RBS-GFP-DT-(1)||Plusgrow medium(+CP)
+|-
+|8/29 Plux-RBS-GFP-DT-(2)||Plusgrow medium(+CP)
+|-
+|8/29 Pbad/araC-RBS-RFP-DT-(1)||Plusgrow medium(+CP)
+|-
+|8/29 Pbad-araC-RBS-RFP-DT-(2)||Plusgrow medium(+CP)
+|-
+|RBS-lysis3-DT-(1)||Plusgrow medium(+CP)
+|}
 </div>

Template:Kyoto/Notebook/Aug 30

From 2013.igem.org

Revision as of 12:15, 22 September 2013

Contents

Aug 30

Gel Extraction

Miniprep

Colony PCR

Ethanol Precipitation

Restriction Enzyme Digestion

Electrophoresis

Liquid Culture