Template:Kyoto/Notebook/Sep 27

From 2013.igem.org

(Difference between revisions)

Revision as of 07:08, 27 September 2013

Sep 27

Gel Extraction

Kojima

Lane	DNA	Enzyme
1	1kbp ladder	--
2	pSB1C3	EcoRI+SpeI
3
4
6	Pcon-attenuator-aptmer-DT	EcoRI+SpeI
7
8
10	Pcon-attenuator-aptmer-DT	EcoRI+SpeI
11
12

Name	concentration[µg/mL]	260/280	260/230
Pcon-attenuator-DT(EcoRI+SpeI)	6.8	1.98	0.43
pT181-attenuator(EcoRI+SpeI)	7.5	1.84	0.06
pSB1C3(EcoRI+SpeI)	17.7	1.97	0.71

Gel Extraction

No name

Lane	DNA	Enzyme
1	1kbp ladder	--
3	pSB4K5	EcoRI+PstI
4	pSB4K5	EcoRI+PstI
5	pSB4K5	EcoRI+PstI
7	pSB6A1	EcoRI+PstI
8	pSB6A1	EcoRI+PstI
9	pSB6A1	EcoRI+PstI

Name	concentration[µg/mL]	260/280	260/230
（´ω｀）
（ΦωΦ^）

</div>

Electrophoresis

No name

Lane	Sample	Enzyme1	Enzyme2
1	pSB4K5	EcoRI	PstI
2	pSB4K5	-	-
3	1kbp ladder	-	-
4	Pcon-attenuator	EcoRI	SpeI
5	Pcon-attenuator	-	-

File:Igku xxxxxx.xxx

Lane	Sample	Enzyme1	Enzyme2
1	RBS-GFP-DT	XbaI	PstI
2	RBS-GFP-DT	-	-
3	RBS-GFP-DT	EcoRI	XbaI
4	1kbp ladder	-	-
5	Ptet	EcoRI	SpeI
6	Ptet	-	-

File:Igku xxxxxx.xxx

Gel Extraction

No name

Lane	DNA	Enzyme
1	100bp ladder	--
3	Ptet	EcoRI+SpeI
4	Ptet	EcoRI+SpeI
5	Ptet	EcoRI+SpeI

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

Lane	DNA	Enzyme
1	1kbp ladder	--
3	Pcon-antisense-aptamer-DT	XbaI+PstI
4	Pcon-antisense-aptamer-DT	XbaI+PstI
5	Pcon-antisense-aptamer-DT	XbaI+PstI

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

Lane	DNA	Enzyme
1	1kbp ladder	--
3	RBS-GFP-DT	EcoRI+XbaI
4	RBS-GFP-DT	EcoRI+XbaI
5	RBS-GFP-DT	EcoRI+XbaI

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

Lane	DNA	Enzyme
1	1kb ladder	--
2	Pcon-attenuator	EcoRI+SpeI
3	Pcon-attenuator	EcoRI+SpeI
4	Pcon-attenuator	EcoRI+SpeI
6	pSB4K5	EcoRI+PstI
7	pSB4K5	EcoRI+PstI
8	pSB4K5	EcoRI+PstI
10	RBS-GFP-DT	XbaI+PstI
11	RBS-GFP-DT	XbaI+PstI
12	RBS-GFP-DT	XbaI+PstI

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

Name	concentration[µg/mL]	260/280	260/230
（´ω｀）
（ΦωΦ^）

Liquid Culture

No name

Sample	medium
Pcon-GFP	Amp
RBS-GFP-DT	CP
Pcon-attenuator	Amp
Pcon-attenuator-aptamer-DT	Amp
Pcon-tetR-DT	Amp

Ligation

No name

state	Vector		Inserter		Ligation High ver.2
experiment	9/17 pSB1C3 (EcoRI+SpeI) 10.0ng/µL	10µL	9/26 pT181 attenuator(EcoRI+SpeI) 7.5ng/µL	1.3µL	3.5 µL
experiment	9/25 DT (EcoRI+XbaI) 42.8ng/µL	2.3µL	9/22 Pcon-pT181 antisense(EcoRI+SpeI) 46.7ng/µL	1.5µL	1.9 µL
experiment	9/27 pSB4K5(EcoRI+PstI) 12.9ng/µL	7.8µL	9/10 Pcon-attenuator(EcoRI+SpeI) 13.8ng/µL	8.0µL	3.5 µL
experiment	9/27 pSB4K5(EcoRI+PstI) 12.9ng/µL	7.8µL	9/25 RBS-GFP-DT(XbaI+PstI) 13.9ng/µL	12µL	3.5 µL
experiment	9/8 pSB4K5 (EcoRI+SpeI) 17.6ng/µL	xx µL	8/21 Pcon-GFP-DT(EcoRI+SpeI) 11ng/µL	12.6µL	xx µL

Liquid Culture

Hirano

Sample	medium
Pcon-spinach-DT	plusgrow Amp
Pcon-tetRaptamer-DT	plusgrow Amp
Pcon-antisense-spinach-DT	plusgrow Amp
spinach-DT	plusgrow CP

RNA Extraction

Nakamoto

Name	concentration[µg/mL]	260/280	260/230
9/26 Pcon-GFP-DT	423.0	1.83	1.46
9/26 Pcon-GFP-DT K	298.7	1.82	1.50

Electrophoresis

No name

Lane	Sample
1	9/27 Pcon-GFP-DT(RNA)
2	1kb ladder
3	9/27 Pcon-GFP-DT K(RNA)

Miniprep

Tatsui

DNA
Pcon-aptamer-DT
Pcon-spinach-DT
Pcon-antisense
Pcon-tetR-DT
Pcon-attenuator-DT
Pcon-antisense-spinach-DT

Transformation

Hirano

Name1	Name2	Sample1(µL)	Sample2(µL)	Competent Cells(µL)	Total(µL)
Pcon-attenuator+RBS-GFP-DT(4K5)	Pcon-attenuator-DT(1A2)	2	1	30	33
Pcon-attenuator+RBS-GFP-DT(4K5)	Pcon-antisense-spinach-DT(1A2)	2	1	30	33
Pcon-attenuator+RBS-GFP-DT(4K5)	Pcon-tetR aptamer-DT(1A2)	2	1	30	33
Pcon-RBS-GFP-DT(4K5)	Pcon-attenuator-DT(1A2)	2	1	30	33
Pcon-RBS-GFP-DT(4K5)	Pcon-antisense-spinach-DT(1A2)	2	1	30	33
Pcon-RBS-GFP-DT(4K5)	Pcon-tetR aptamer-DT(1A2)	2	1	30	33
Pcon-attenuator+RBS-GFP-DT(4K5)	--	2	--	20	22
Pcon-RBS-GFP-DT(4K5)	--	2	--	20	22
attenuator(1C3)	--	2	--	20	22

Plating

Hirano

Sample	Use plate
9/26 GGA2	LB(CP)
9/26 GGA3	LB(CP)
9/26 GGA4	LB(CP)
9/26 GGA5	LB(CP)
9/26 GGA6	LB(CP)
9/26 GGA7	LB(CP)
9/26 GGA11	LB(CP)
9/26 GGA312	LB(CP)
9/26 GGA13	LB(CP)
9/26 GGA14	LB(CP)
9/26 GGA15	LB(CP)
9/26 GGA20	LB(CP)
9/26 GGA23	LB(CP)
9/26 GGA24	LB(CP)
9/26 Ptet+RBS-GFP-DT	LB(CP)

incubate 37°C

Restriction Enzyme Digestion

Kojima

	9/24 pSB1C3	EcoRI	SpeI	buffer	MilliQ	total
2cuts	11µL	1µL	1µL	3µL	14µL	30µL
NC	0.5µL	0µL	0µL	1µL	8.5µL	10µL

	9/24 Pcon-pT181attenuator-aptamer12-1R-DT	EcoRI	SpeI	buffer	MilliQ	total
2cuts	8.7µL	1µL	1µL	3µL	16.3µL	30µL
NC	0.4µL	0µL	0µL	1µL	8.6µL	10µL

	9/24 pT181attenuator	EcoRI	SpeI	buffer	MilliQ	total
2cuts	3.1µL	1µL	1µL	3µL	21.9µL	30µL
NC	0.2µL	0µL	0µL	1µL	8.8µL	10µL

	8/17 RBS-GFP-DT	EcoRI	BSA	buffer	MilliQ	total
1cut	16.4µL	1µL	3µL	3µL	6.6µL	30µL
NC	0.4µL	0µL	1µL	1µL	7.6µL	10µL

	9/17 pSB4K5	EcoRI	PstI	BSA	buffer	MilliQ	total
2cuts	8.5µL	1µL	1µL	3µL	3µL	13.5µL	30µL
NC	0.4µL	0µL	0µL	1µL	1µL	7.6µL	10µL

	9/14 pSB4K5	EcoRI	PstI	BSA	buffer	MilliQ	total
2cuts	7.3µL	1µL	1µL	3µL	3µL	14.7µL	30µL
NC	0.4µL	0µL	0µL	1µL	1µL	7.6µL	10µL

	9/22 Pcon-pT181attenuator-DT	EcoRI	SpeI	buffer	MilliQ	total
2cuts	5.4µL	1µL	1µL	3µL	19.6µL	30µL
NC	0.3µL	0µL	0µL	1µL	8.7µL	10µL

	8/7 RBS-GFP-DT 1	XbaI	PstI	BSA	buffer	MilliQ	total
2cuts	8.1µL	1µL	1µL	3µL	3µL	13.9µL	30µL
NC	0.4µL	0µL	0µL	1µL	1µL	7.6µL	10µL

	9/26 RBS-GFP-DT 1	XbaI	BSA	buffer	MilliQ	total
1cut	8.1µL	1µL	3µL	3µL	14.9µL	30µL

	9/16 Pλ-luxI	EcoRI	PstI	BSA	buffer	MilliQ	total
2cuts	14.7µL	1µL	1µL	3µL	3µL	7.3µL	30µL
NC	0.7µL	0µL	0µL	1µL	1µL	7.3µL	10µL

	9/21 Ptet	EcoRI	SpeI	buffer	MilliQ	total
2cuts	14.2µL	1µL	1µL	3µL	10.8µL	30µL
NC	0.7µL	0µL	0µL	1µL	8.3µL	10µL

	9/24 Pcon-pT181attenuator-aptamer12-1R-DT	XbaI	PstI	BSA	buffer	MilliQ	total
2cuts	8.7µL	1µL	1µL	3µL	3µL	13.3µL	30µL
NC	0.4µL	0µL	0µL	1µL	1µL	7.6µL	10µL

Electrophoresis

Kojima

Lane	Sample	Enzyme1	Enzyme2
1	pSB1C3	EcoRI	SpeI
2	pSB1C3	--	--
3	Pcon-pT181attenuator-aptamer12-1R-DT	EcoRI	SpeI
4	Pcon-pT181attenuator-aptamer12-1R-DT	--	--
5	1kb ladder	--	--
6	pT181attenuator	EcoRI	SpeI
7	pT181attenuator	--	--
8	RBS-GFP-DT	EcoRI	--
9	RBS-GFP-DT	--	--

File:IgkuxxxxBa 1.jpg

@@ Line 312: / Line 312: @@
 |}
 </div>
+===Plating===
+<div class="experiment">
+<span class="author">Hirano</span>
+</div>
+{| class="wikitable"
+!Sample||Use plate
+|-
+|9/26 GGA2 ||LB(CP)
+|-
+|9/26 GGA3 ||LB(CP)
+|-
+|9/26 GGA4||LB(CP)
+|-
+|9/26 GGA5 ||LB(CP)
+|-
+|9/26 GGA6 ||LB(CP)
+|-
+|9/26 GGA7 ||LB(CP)
+|-
+|9/26 GGA11 ||LB(CP)
+|-
+|9/26 GGA312||LB(CP)
+|-
+|9/26 GGA13 ||LB(CP)
+|-
+|9/26 GGA14||LB(CP)
+|-
+|9/26 GGA15 ||LB(CP)
+|-
+|9/26 GGA20||LB(CP)
+|-
+|9/26 GGA23 ||LB(CP)
+|-
+|9/26 GGA24||LB(CP)
+|-
+|9/26 Ptet+RBS-GFP-DT ||LB(CP)
+|}
+</div>
+incubate 37&deg;C
+===Restriction Enzyme Digestion===
+<div class="experiment">
+<span class="author">Kojima</span>
+{| class="wikitable"
+!||9/24 pSB1C3||EcoRI||SpeI||buffer||MilliQ||total
+|-
+|2cuts||11&micro;L||1&micro;L||1&micro;L||3&micro;L||14&micro;L||30&micro;L
+|-
+|NC||0.5&micro;L||0&micro;L||0&micro;L||1&micro;L||8.5&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||9/24 Pcon-pT181attenuator-aptamer12-1R-DT||EcoRI||SpeI||buffer||MilliQ||total
+|-
+|2cuts||8.7&micro;L||1&micro;L||1&micro;L||3&micro;L||16.3&micro;L||30&micro;L
+|-
+|NC||0.4&micro;L||0&micro;L||0&micro;L||1&micro;L||8.6&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||9/24 pT181attenuator||EcoRI||SpeI||buffer||MilliQ||total
+|-
+|2cuts||3.1&micro;L||1&micro;L||1&micro;L||3&micro;L||21.9&micro;L||30&micro;L
+|-
+|NC||0.2&micro;L||0&micro;L||0&micro;L||1&micro;L||8.8&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||8/17 RBS-GFP-DT||EcoRI||BSA ||buffer||MilliQ||total
+|-
+|1cut||16.4&micro;L||1&micro;L||3&micro;L||3&micro;L||6.6&micro;L||30&micro;L
+|-
+|NC||0.4&micro;L||0&micro;L||1&micro;L||1&micro;L||7.6&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||9/17 pSB4K5||EcoRI||PstI||BSA ||buffer||MilliQ||total
+|-
+|2cuts||8.5&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||13.5&micro;L||30&micro;L
+|-
+|NC||0.4&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.6&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||9/14 pSB4K5||EcoRI||PstI||BSA ||buffer||MilliQ||total
+|-
+|2cuts||7.3&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.7&micro;L||30&micro;L
+|-
+|NC||0.4&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.6&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||9/22 Pcon-pT181attenuator-DT||EcoRI||SpeI ||buffer||MilliQ||total
+|-
+|2cuts||5.4&micro;L||1&micro;L||1&micro;L||3&micro;L||19.6&micro;L||30&micro;L
+|-
+|NC||0.3&micro;L||0&micro;L||0&micro;L||1&micro;L||8.7&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||8/7 RBS-GFP-DT 1||XbaI||PstI||BSA ||buffer||MilliQ||total
+|-
+|2cuts||8.1&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||13.9&micro;L||30&micro;L
+|-
+|NC||0.4&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.6&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||9/26 RBS-GFP-DT 1||XbaI ||BSA ||buffer||MilliQ||total
+|-
+|1cut||8.1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.9&micro;L||30&micro;L
+|}
+{| class="wikitable"
+!||9/16 P&lambda;-luxI||EcoRI||PstI||BSA ||buffer||MilliQ||total
+|-
+|2cuts||14.7&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||7.3&micro;L||30&micro;L
+|-
+|NC||0.7&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.3&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||9/21 Ptet||EcoRI||SpeI ||buffer||MilliQ||total
+|-
+|2cuts||14.2&micro;L||1&micro;L||1&micro;L||3&micro;L||10.8&micro;L||30&micro;L
+|-
+|NC||0.7&micro;L||0&micro;L||0&micro;L||1&micro;L||8.3&micro;L||10&micro;L
+|}
+{| class="wikitable"
+!||9/24 Pcon-pT181attenuator-aptamer12-1R-DT||XbaI||PstI||BSA ||buffer||MilliQ||total
+|-
+|2cuts||8.7&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||13.3&micro;L||30&micro;L
+|-
+|NC||0.4&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.6&micro;L||10&micro;L
+|}
+</div>
+===Electrophoresis===
+<div class="experiment">
+<span class="author">Kojima</span>
+{| class="wikitable"
+!Lane||Sample|| Enzyme1|| Enzyme2
+|-
+|1||pSB1C3||EcoRI||SpeI
+|-
+|2||pSB1C3||--||--
+|-
+|3||Pcon-pT181attenuator-aptamer12-1R-DT||EcoRI||SpeI
+|-
+|4||Pcon-pT181attenuator-aptamer12-1R-DT||--||--
+|-
+|5||1kb ladder||--||--
+|-
+|6||pT181attenuator||EcoRI||SpeI
+|-
+|7||pT181attenuator||--||--
+|-
+|8||RBS-GFP-DT||EcoRI||--
+|-
+|9||RBS-GFP-DT||--||--
+|}
+[[File:IgkuxxxxBa 1.jpg]]<br>

Template:Kyoto/Notebook/Sep 27

From 2013.igem.org

Revision as of 07:08, 27 September 2013

Contents

Sep 27

Gel Extraction

Gel Extraction

Electrophoresis

Gel Extraction

Liquid Culture

Ligation

Liquid Culture

RNA Extraction

Electrophoresis

Miniprep

Transformation

Plating

Restriction Enzyme Digestion

Electrophoresis