Template:Kyoto/Notebook/Aug 30

From 2013.igem.org

(Difference between revisions)

Latest revision as of 03:49, 28 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	--	--

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	--	--

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)

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 30: / Line 30: @@
 !DNA||concentration[&micro;g/mL]||260/280||260/230
 |-
-|RBS-lysis2 9||264.6　||1.97　||2.21
+|RBS-lysis2 9||264.6||1.97||2.21
 |-
-|RBS-lysis2 12||220.0　||1.89　||2.46
+|RBS-lysis2 12||220.0||1.89||2.46
 |-
-|RBS-lysis2 14||195.3　||1.88　||1.53
+|RBS-lysis2 14||195.3||1.88||1.53
 |}
 </div>
 ===Colony PCR===
 <div class="experiment">
@@ Line 58: / Line 59: @@
 |5min||30s||30s||2min||30cycle
 |}
+[[File:Igku_Aug30_colonyPCR_N1.jpg]]
 </div>
@@ Line 179: / Line 181: @@
 |14||100bp ladder||--||--
 |}
-[[File:igku_Aug30electrophoresis1]]<br>
+[[File:Igku Aug30 Electrophoresis(N2-1).jpg]]<br>
@@ Line 201: / Line 203: @@
 |8||100bp ladder||--||--
 |}
-[[File:igku_Aug30electrophoresis2]]<br>
+[[File:Igku Aug30 Electrophoresis(N2-2).jpg]]<br>
 {| class="wikitable"
@@ Line 208: / Line 210: @@
 |1||1kb ladder
 |-
-|2||8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)
+|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 Aug30 Electrophoresis(N2-3).jpg]]<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)
 |-
-|3||8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)
+|8/29 Plux-RBS-GFP-DT-(1)||Plusgrow medium(+CP)
 |-
-|4||8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)
+|8/29 Plux-RBS-GFP-DT-(2)||Plusgrow medium(+CP)
 |-
-|5||8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)
+|8/29 Pbad/araC-RBS-RFP-DT-(1)||Plusgrow medium(+CP)
 |-
-|6||Pbad/araC-RBS-RFP-DT-(1)
+|8/29 Pbad-araC-RBS-RFP-DT-(2)||Plusgrow medium(+CP)
 |-
-|7||Pbad-araC-RBS-RFP-DT-(2)
+|RBS-lysis3-DT-(1)||Plusgrow medium(+CP)
 |}
-[[File:igku_Aug30electrophoresis3]]<br>
 </div>

Template:Kyoto/Notebook/Aug 30

From 2013.igem.org

Latest revision as of 03:49, 28 September 2013

Contents

Aug 30

Gel Extraction

Miniprep

Colony PCR

Ethanol Precipitation

Restriction Enzyme Digestion

Electrophoresis

Liquid Culture