Template:Kyoto/Notebook/Sep 14

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Sep 14

Hirano

	9/14 pSB4K5S	EcoRI	XbaI	SpeI	PstI	BufferB	BufferD	BSA	MilliQ	total
2cuts	10µL	1µL	0µL	1µL	0µL	3µL	0µL	3µL	12µL	30µL
NC	0.5µL	0µL	0µL	0µL	0µL	1µL	0µL	1µL	7.5µL	10µL

	9/13 Plac	XbaI	PstI	BufferH	BSA	MilliQ	total
1 cut	9µL	0µL	1µL	3µL	3µL	14µL	30µL
NC	1µL	0µL	0µL	1µL	1µL	7µL	10µL

	9/14 PKaiBC	EcoRI	XbaI	SpeI	PstI	BufferB	BSA	MilliQ	total
2cuts	11µL	1µL	0µL	1µL	0µL	3µL	3µL	11µL	30µL
NC	4µL	0µL	0µL	0µL	0µL	1µL	1µL	4µL	10µL

	9/14 RpaB	EcoRI	XbaI	SpeI	PstI	BufferD	BSA	MilliQ	total
2cuts	13µL	0µL	1µL	0µL	1µL	3µL	3µL	9µL	30µL
NC	2µL	0µL	0µL	0µL	0µL	1µL	1µL	5µL	10µL

	8/17 RBS-GFP-DT	EcoRI	XbaI	SpeI	PstI	BufferD	BSA	MilliQ	total
1cut	9µL	0µL	1µL	0µL	0µL	3µL	3µL	14µL	30µL
NC	0.5µL	0µL	0µL	0µL	0µL	1µL	1µL	7.5µL	10µL

Hirano

Sample	medium
9/11 Plac(BBa-R0011)-(3)	Plusgrow medium(+Amp)

No name

genome DNA	MilliQ	Big Dye	5x buffer	temp(400mg)	primer	total
Pcon-RBS-luxR-DT	0.5	1.75	1.75	0.4	0.5	5.1	10.5
Pcon-RBS-luxR-DT	0.5	1.75	1.75	0.4	0.5	5.1	10.5
Pcon-RBS-tetR-DT	0.5	1.75	1.75	3	0.5	3	10.5
Pcon-RBS-tetR-DT	0.5	1.75	1.75	3	0.5	3	10.5
Pcon-RBS-luxR-DT	0.5	1.75	1.75	2.4	0.5	3.6	10.5
Plux-RBS-GFP-DT	0.5	1.75	1.75	1.9	0.5	4.1	10.5
Plux-RBS-GFP-DT	0.5	1.75	0.5	1.9	0.5	4.1	10.5
Pcon-RBS-GFP-DT	0.5	1.75	0.5	1.2	0.5	4.8	10.5
RBS-lysis3-DT	0.5	1.75	0.5	1.0	0.5	5.0	10.5
Pcon-RBS-GFP-DT	0.5	1.75	0.5	1.4	0.5	4.6	10.5
Pcon-RBS-GFP-DT	0.5	1.75	0.5	1.4	0.5	4.6	10.5
RBS-lysis3-DT	0.5	1.75	1.75	0.4	0.5	5.1	10.5
Ptet-RBS-spinach-DT	0.5	1.75	0.5	0.59	0.5	0.1	10.5

PreDenature	Denature	Annealing	Extension	cycle
96°C	96°C	50°C	60°C	--
2min	10s	5s	4min	30cycle

No name

	9/14 Plac(1A2)	EcoRI	XbaI	SpeI	PstI	BufferB	BSA	MilliQ	total
2cuts	23µL	0µL	0µL	0.3µL	0µL	3µL	3µL	0µL	30µL

	9/14 RBS-GFP-DT	EcoRI	XbaI	SpeI	PstI	BufferH	BSA	MilliQ	total
2 cuts	23µL	1µL	0µL	0µL	0µL	3µL	3µL	0µL	30µL

Nakamoto and Hirano

state	Vector		Inserter		Ligation High ver.2
experiment	8/30 pSB1C3 (EcoRI & SpeI)	6.6	8/30 tRNA-Spinach (EcoRI & SpeI)	12.5	3.5
experiment	8/30 pSB1C3 (EcoRI & SpeI)	6.6	8/30 pT181 antisense (EcoRI & SpeI)	12.5	3.5
experiment	8/30 pSB1C3 (XbaI & Pst)	12.5	8/30 pT181 attenuator (XbaI & PstI)	25	3.5
experiment	8/30 pSB1C3 (XbaI & Pst)	12.5	8/30 pT181 antisense (XbaI & PstI)	25	3.5
experiment	8/20 DT (EcoRI & XbaI)	6.6	8/30 pT181 attenuator(2) (EcoRI & SpeI)	12.5	3.5
experiment	8/20 DT (EcoRI & XbaI)	6.6	8/30 pT181 antisense (EcoRI & SpeI)	12.5	3.5
experiment	8/27 DT (EcoRI & XbaI)	3.0	8/30 tRNA-Spinach (EcoRI & SpeI)	12.5	3.5
experiment	8/20 DT (EcoRI & XbaI)	6.6	8/30 RBS-lysis2 (EcoRI & SpeI)	12.5	3.5
experiment	8/22 J23100 (SpeI & PstI)	--	RBS-tetR-DT (XbaI & PstI)	--	3.5
experiment	8/22 Plac (SpeI & PstI)	8.0	8/18 RBS-lacZα-DT (XbaI & PstI)	10	3.5

Samples were evaporeted used evaporator into about 7 µL

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

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)

No Name

Name	Sample	Competent Cells	Total	Plate
8/27 pT181 attenuator-DT	2µL	20µL	22µL	CP
8/27 RBS-lysis3-DT	2µL	20µL	22µL	CP
8/28 RBS-lysis3-DT	2µL	20µL	22µL	CP
8/28 Pcon-RBS-GFP-DT-Pcon-RBS-LuxR-DT	2µL	20µL	22µL	Amp
8/28 RBS-lysis1-DT	2µL	20µL	22µL	CP
8/28 Plux-RBS-GFP-DT	2µL	20µL	22µL	CP
8/28 Spinach-DT	2µL	20µL	22µL	CP