Template:Kyoto/Notebook/Aug 22

From 2013.igem.org

(Difference between revisions)

Latest revision as of 14:49, 25 September 2013

Aug 22

Liquid culture

Nakamoto

Sample	Medium
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (1)	8/21 Plusgrow medium (+Amp)
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (2)	8/21 Plusgrow medium (+Amp)
8/21 Pcon-RBS-GFP-DT control (1)	8/21 Plusgrow medium (+Amp)
8/21 Pcon-RBS-GFP-DT control (2)	8/21 Plusgrow medium (+Amp)
8/21 RBS-lysis1 (1)	8/21 Plusgrow medium (+Amp)
8/21 RBS-lysis1 (2)	8/21 Plusgrow medium (+Amp)
8/21 RBS control (1)	8/21 Plusgrow medium (+Amp)
8/21 RBS control (2)	8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm)-RBS-tetR-DT (1)	8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm)-RBS-tetR-DT (2)	8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm) control (1)	8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm) control (2)	8/21 Plusgrow medium (+Amp)
8/21 Plux-RBS-GFP-DT (1)	8/21 Plusgrow medium (+Amp)
8/21 pSB1C3(BBa_J04450) (1)	8/21 Plusgrow medium (+Amp)
8/21 pSB1C3(BBa_J04450) (2)	8/21 Plusgrow medium (+Amp)

incubate 37°C 10hour

Colony PCR

Kojima

Sample	base pair
8/21 RBS-lysis1(1)	400
8/21 RBS-lysis1(2)	400
8/21 RBS control(1)	--
8/21 RBS control(2)	--
negative control	--

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

Sample	base pair
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (1)	2143
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (2)	2143
8/21 Pcon-RBS-luxR control(1)	--
8/21 Pcon-RBS-luxR control(2)	--
negative control	--

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

Sample	base pair
8/21 Ptet-RBS-tetR-DT (1)	1216
8/21 Ptet-RBS-tetR-DT (2)	1216
8/21 Ptet control (1)	--
8/21 Ptet control (2)	--
8/21 Plux-RBS-GFP-DT (1)	1227
8/21 pSB1C3(BBa_J04450) (1)	1353
8/21 pSB1C3(BBa_J04450) (2)	1353
negative control	--

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

Restriction Enzyme Digestion

Tatsui, Kojima, Honda and Ashida

	Fusion1 antisense (1) (300 µg/mL)	EcoRI	SpeI	buffer H	MilliQ	total
2 cuts	3.3	1	1	3	21.7	30
1 cut	0.7	0.2	0	1	8.1	10
1 cut	0.7	0	0.2	1	8.1	10
NC	0.7	0	0	1	8.3	10

	Fusion1 antisense (1) (300 µg/mL)	XbaI	PstI	buffer M	BSA	MilliQ	total
2 cuts	3.3	1	1	3	3	18.7	30
1 cut	0.7	0.2	0	1	1	7.1	10
1 cut	0.7	0	0.2	1	1	7.1	10
NC	0.7	0	0	1	1	7.3	10

	Fusion6 antisense (1) (336 µg/mL)	EcoRI	SpeI	buffer H	MilliQ	total
2 cuts	3	1	1	3	22	30
1 cut	0.6	0.2	0	1	8.2	10
1 cut	0.6	0	0.2	1	8.2	10
NC	0.6	0	0	1	8.4	10

	Fusion6 antisense (1) (336 µg/mL)	XbaI	PstI	buffer M	BSA	MilliQ	total
2 cuts	3	1	1	3	3	19	30
1 cut	0.6	0.2	0	1	1	7.2	10
1 cut	0.6	0	0.2	1	1	7.2	10
NC	0.6	0	0	1	1	7.4	10

	pT181 antisense (1) (178 µg/mL)	EcoRI	SpeI	buffer H	MilliQ	total
2 cuts	5.6	1	1	3	19.4	30
1 cut	1.1	0.2	0	1	7.7	10
1 cut	1.1	0	0.2	1	7.7	10
NC	1.1	0	0	1	7.9	10

	pT181 antisense (1) (178 µg/mL)	XbaI	PstI	buffer M	BSA	MilliQ	total
2 cuts	5.6	1	1	3	3	16.4	30
1 cut	1.1	0.2	0	1	1	6.7	10
1 cut	1.1	0	0.2	1	1	6.7	10
NC	1.1	0	0	1	1	6.9	10

	pT181 attenuator (1)	EcoRI	SpeI	buffer H	MilliQ	total
2 cuts	3	1	1	3	22	30
1 cut	0.6	0.2	0	1	8.2	10
1 cut	0.6	0	0.2	1	8.2	10
NC	0.6	0	0	1	8.4	10

	pT181 attenuator (1)	XbaI	PstI	buffer M	BSA	MilliQ	total
2 cuts	3	1	1	3	3	19	30
1 cut	0.6	0.2	0	1	1	7.2	10
1 cut	0.6	0	0.2	1	1	7.2	10
NC	0.6	0	0	1	1	7.4	10

	Fusion1 attenuator (1)	EcoRI	SpeI	buffer H	MilliQ	total
2 cuts	3.3	1	1	3	21.7	30
1 cut	0.7	0.2	0	1	8.1	10
1 cut	0.7	0	0.2	1	8.1	10
NC	0.7	0	0	1	8.3	10

	Fusion1 attenuator (1)	XbaI	PstI	buffer M	BSA	MilliQ	total
2 cuts	3.3	1	1	3	3	18.7	30
1 cut	0.7	0.2	0	1	1	7.1	10
1 cut	0.7	0	0.2	1	1	7.1	10
NC	0.7	0	0	1	1	7.3	10

	Fusion3m2 attenuator (1)	EcoRI	SpeI	buffer H	MilliQ	total
2 cuts	3	1	1	3	22	30
1 cut	0.6	0.2	0	1	8.2	10
1 cut	0.6	0	0.2	1	8.2	10
NC	0.6	0	0	1	8.4	10

	Fusion3m2 attenuator (1)	XbaI	PstI	buffer M	BSA	MilliQ	total
2 cuts	3	1	1	3	3	19	30
1 cut	0.6	0.2	0	1	1	7.2	10
1 cut	0.6	0	0.2	1	1	7.2	10
NC	0.6	0	0	1	1	7.4	10

	Spinach (2)	EcoRI	SpeI	buffer B	BSA	MilliQ	total
2 cuts	2.3	0.5	0.5	3	0.3	23.4	30
1 cut	0.5	0.1	0	1	0.1	8.3	10
1 cut	0.5	0	0.1	1	0.1	8.3	10
NC	0.5	0	0	1	0.1	8.4	10

	tetR aptamer 12_P (2)	EcoRI	SpeI	buffer B	BSA	MilliQ	total
2 cuts	2.7	0.5	0.5	3	0.3	23	30
1 cut	0.5	0.1	0	1	0.1	8.3	10
1 cut	0.5	0	0.1	1	0.1	8.3	10
NC	0.5	0	0	1	0.1	8.4	10

	tetR aptamer 12_1R (2)	EcoRI	SpeI	buffer B	BSA	MilliQ	total
2 cuts	2.6	0.5	0.5	3	0.3	23.1	30
1 cut	0.5	0.1	0	1	0.1	8.3	10
1 cut	0.5	0	0.1	1	0.1	8.3	10
NC	0.5	0	0	1	0.1	8.4	10

	tetR aptamer 12_1M (1)	EcoRI	SpeI	buffer B	BSA	MilliQ	total
2 cuts	2.7	0.5	0.5	3	0.3	23	30
1 cut	0.5	0.1	0	1	0.1	8.3	10
1 cut	0.5	0	0.1	1	0.1	8.3	10
NC	0.5	0	0	1	0.1	8.4	10

	J23100	SpeI	PstI	buffer B	BSA	MilliQ	total
2 cuts	7	0.5	0.5	3	0.3	18.7	30
1 cut	1.4	0.1	0	1	0.1	7.4	10
1 cut	1.4	0	0.1	1	0.1	7.4	10
NC	1.4	0	0	1	0.1	7.5	10

	Plac (2)	SpeI	PstI	buffer B	BSA	MilliQ	total
2 cuts	11.2	0.5	0.5	3	0.3	14.5	30
1 cut	2.2	0.1	0	1	0.1	6.6	10
1 cut	2.2	0	0.1	1	0.1	6.6	10
NC	2.2	0	0	1	0.1	6.7	10

Electrophoresis

No name

Lane	Sample
1	100bp ladder
2	RBS-lysis1 -(1)
3	RBS-lysis1 -(2)
4	RBS-lysis2 -(1)
5	RBS-lysis2 -(2)
6	RBS(NC) -(1)
7	RBS(NC) -(2)
8	NC

Lane	Sample
1	1kb ladder
2	Ptet-RBS-tetR-DT -(1)
3	Ptet-RBS-tetR-DT -(2)
4	Ptet(NC) -(1)
5	Ptet(NC) -(2)
6	Plux-RBS-GFP-DT -(1)
7	pSB1C3 -(1)
8	pSB1C3 -(2)
9	NC

Lane	Sample
1	1kb ladder
2	Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT -(1)
3	Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT -(2)
4	Pcon-RBS-luxR -(1)
5	Pcon-RBS-luxR -(2)
6	NC
7	1kb ladder

Miniprep

Ashida

DNA	concentration[µg/mL]	260/280	260/230
8/22 RBS-lysis (1)	128	1.77	2.29
8/22 RBS-lysis (2)	96	1.80	2.07
8/22 pSB1C3 (1)	108	1.55	1.45
8/22 pSB1C3 (2)	130	1.69	1.76

Electrophoresis

No name

Lane	Sample	Enzyme1	Enzyme2
1	100bp	--	--
2	8/22 pT181 attenuator (1)	EcoRI	SpeI
3	8/22 pT181 attenuator (1)	EcoRI	--
4	8/22 pT181 attenuator (1)	--	SpeI
5	8/22 pT181 attenuator (1)	--	--
6	8/22 pT181 attenuator (1)	XbaI	PstI
7	8/22 pT181 attenuator (1)	XbaI	--
8	8/22 pT181 attenuator (1)	--	PstI
9	8/22 pT181 attenuator (1)	--	--
10	8/22 Fusion6 antisense (1)	EcoRI	SpeI
11	8/22 Fusion6 antisense (1)	EcoRI	--
12	8/22 Fusion6 antisense (1)	--	SpeI
13	8/22 Fusion6 antisense (1)	--	--
14	100bp	--	--

Lane	Sample	Enzyme1	Enzyme2
1	100bp	--	--
2	8/22 Fusion6 antisense (1)	XbaI	PstI
3	8/22 Fusion6 antisense (1)	XbaI	--
4	8/22 Fusion6 antisense (1)	--	PstI
5	8/22 Fusion6 antisense (1)	--	--
6	8/22 Fusion1 antisense (1)	EcoRI	SpeI
7	8/22 Fusion1 antisense (1)	EcoRI	--
8	8/22 Fusion1 antisense (1)	--	SpeI
9	8/22 Fusion1 antisense (1)	--	--
10	8/22 Fusion1 antisense (1)	XbaI	PstI
11	8/22 Fusion1 antisense (1)	XbaI	--
12	8/22 Fusion1 antisense (1)	--	PstI
13	8/22 Fusion1 antisense (1)	--	--
14	100bp	--	--

@@ Line 313: / Line 313: @@
 |NC||2.2||0||0||1||0.1||6.7||10
 |}
+</div>
+===Electrophoresis===
+<div class="experiment">
+<span class="author">No name</span>
+{| class="wikitable"
+!Lane||Sample
+|-
+|1||100bp ladder
+|-
+|2||RBS-lysis1 -(1)
+|-
+|3||RBS-lysis1 -(2)
+|-
+|4||RBS-lysis2 -(1)
+|-
+|5||RBS-lysis2 -(2)
+|-
+|6||RBS(NC) -(1)
+|-
+|7||RBS(NC) -(2)
+|-
+|8||NC
+|}
+[[File:Igku Aug22 Electrophoresis(ColoP)-1.jpg]]<br>
+{| class="wikitable"
+!Lane||Sample
+|-
+|1||1kb ladder
+|-
+|2||Ptet-RBS-tetR-DT -(1)
+|-
+|3||Ptet-RBS-tetR-DT -(2)
+|-
+|4||Ptet(NC) -(1)
+|-
+|5||Ptet(NC) -(2)
+|-
+|6||Plux-RBS-GFP-DT -(1)
+|-
+|7||pSB1C3 -(1)
+|-
+|8||pSB1C3 -(2)
+|-
+|9||NC
+|}
+[[File:Igku Aug22 Electrophoresis(ColoP)-2.jpg]]<br>
+{| class="wikitable"
+!Lane||Sample
+|-
+|1||1kb ladder
+|-
+|2||Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT -(1)
+|-
+|3||Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT -(2)
+|-
+|4||Pcon-RBS-luxR -(1)
+|-
+|5||Pcon-RBS-luxR -(2)
+|-
+|6||NC
+|-
+|7||1kb ladder
+|}
+[[File:Igku Aug22 Electrophoresis(ColoP)-3.jpg]]<br>
+</div>
+===Miniprep===
+<div class="experiment">
+<span class="author">Ashida</span>
+{|class="wikitable"
+!DNA||concentration[&micro;g/mL]||260/280||260/230
+|-
+|8/22 RBS-lysis (1)||128||1.77||2.29
+|-
+|8/22 RBS-lysis (2)||96||1.80||2.07
+|-
+|8/22 pSB1C3 (1)||108||1.55||1.45
+|-
+|8/22 pSB1C3 (2)||130||1.69||1.76
+|}
 </div>

Template:Kyoto/Notebook/Aug 22

From 2013.igem.org

Latest revision as of 14:49, 25 September 2013

Contents

Aug 22

Liquid culture

Colony PCR

Restriction Enzyme Digestion

Electrophoresis

Miniprep

Electrophoresis