Template:Kyoto/Notebook/Sep 15

From 2013.igem.org

Sep 15

Miniprep

No name

DNA	concentration[µg/mL]	260/280	260/230
Pλ-luxI	225	1.59	0.83

Liquid Culture

No name

Sample	medium
9/13 Pλ-luxI-1
9/7 pSB4K5-1
9/11 Plac-3

Ligation

No name

state	Vector		Inserter		Ligation High ver.2
experiment	9/13 Pcon(SpeI & PstI)	3.6 µL	9/12 RBS-lacZα-DT(XbaI & PstI)	4.7 µL	8 µL

Restriction Enzyme Digestion

No Name

	DNA	EcorI	SpeI	Buffer	BSA	MilliQ	total
9/7Pcon-RBS-tetR-DT (EcoRI & SpeI)	15 µL	1 µL	1 µL	3 µL	3 µL	6 µL	30 µL
NC	0.7 µL	0 µL	0 µL	1 µL	1 µL	7.3 µL	10 µL

	DNA	EcoRI	SpeI	Buffer	BSA	MilliQ	total
9/12 Pcon-pT181 attenuator-DT (EcoRI & SpeI)	6.8 µL	1 µL	1 µL	3 µL	3 µL	15.2 µL	30 µL
NC	0.3 µL	0 µL	0 µL	1 µL	1 µL	7.7 µL	10 µL

	DNA	EcoRI	SpeI	Buffer	BSA	MilliQ	total
9/12 Pcon-apt12_1R-DT(EcoRI & SpeI)	6.6 µL	1 µL	1 µL	3 µL	3 µL	15.4 µL	30 µL
NC	0.3 µL	0 µL	0 µL	1 µL	1 µL	7.7 µL	10 µL

	DNA	EcoRI	SpeI	Buffer	BSA	MilliQ	total
9/8 Pcon-spinach-DT (EcoRI & SpeI)	7.6 µL	1 µL	1 µL	3 µL	3 µL	15.4 µL	30 µL
NC	0.3 µL	0 µL	0 µL	1 µL	1 µL	7.7 µL	10 µL

	DNA	XbaI	PstI	Buffer	BSA	MilliQ	total
9/11 Pcon-pT181 anticodone-spinach-DT(XbaI & PstI)	7.2 µL	1 µL	1 µL	3 µL	3 µL	14.8 µL	30 µL
NC	0.4 µL	0 µL	0 µL	1 µL	1 µL	7.6 µL	10 µL

	DNA	XbaI	PstI	Buffer	BSA	MilliQ	total
8/17 RBS-lacZα-DT (XbaI & PstI)	5.3 µL	1 µL	1 µL	3 µL	3 µL	16.7 µL	30 µL
NC	0.5 µL	0 µL	0 µL	1 µL	1 µL	7.5 µL	10 µL

	DNA	EcoRI	XbaI	Buffer	BSA	MilliQ	total
9/7 Ptet-RBS-LacZα-DT (EcoRI & XbaI)	2.9 µL	1 µL	1 µL	3 µL	3 µL	18.5 µL	30 µL
NC	1.5 µL	0 µL	0 µL	1 µL	1 µL	7 µL	10 µL

Electrophoresis

No Name

Lane	Sample	Enzyme1	Enzyme2
1,14	100bp ladder
2	Pcon-RBS-tetR-DT	EcoRI	SpeI
3	Pcon-RBS-tetR-DT
4	Pcon-pT181 attenuator-DT	EcoRI	SpeI
5	Pcon-pT181 attenuator-DT
6	Pcon-apt12-1R-DT	EcoRI	SpeI
7	Pcon-apt12-1R-DT
8	Pcon-spinach-DT	EcoRI	SpeI
9	Pcon-spinach-DT
10	Pcon-pT181 antisense-spinach-DT	XbaI	PstI
11	Pcon-pT181 antisense-spinach-DT
12	RBS-lacZα-DT	XbaI	PstI
13	RBS-lacZα-DT
15	Ptet-RBS-lacZα-DT	EcoRI	XbaI
16	Ptet-RBS-lacZα-DT
17	1kbp ladder

Gel Extraction

No Name 1

Lane	DNA	Enzyme
1	100bp ladder
2~4	Pcon-pT181 attenuator-DT	EcoRI & SpeI
6~8	Pcon-apt12_1R-DT
10~12	Pcon-spinach-DT

2

Lane	DNA	Enzyme
1	100bp ladder
2~4	Pcon-pT181 anticodone-spinach-DT	XbaI & PstI
6~8	RBS-lacZα-DT	XbaI & PstI
10~12	Pcon-RBS-tetR-DT	EcoRI & SpeI

Name	concentration[µg/mL]	260/280	260/230
9/15 Ptet-RBS-lacZα-DT (EcoRI & SpeI)	1.9	1.08	0.77
9/15 Pcon-RBS-tetR-DT (EcoRI & SpeI)	5.9	1.30	0.17
9/15 RBS-lacZα-DT (XbaI & PstI)	1.0	0.85	0.44
Pcon-pT181 anticodone-spinach-DT (XbaI & PstI)	1.3	0.95	0.28
Pcon-spinach-DT	0.9	1.37	0.05
Pcon-apt12_1R-DT	1.7	1.42	0.29
Pcon-pT181 attenuator-DT (EcoRI & SpeI)	2.1	1.53	0.46

Ligation

No Name

state	Vector		Inserter		Ligation High ver.2
experiment	9/13 pSB1C3 EcoRI & SpeI	4.7 µL	9/15 Pcon-RBS-tetR-DT EcoRI & SpeI	1.9 µL	xx µL
experiment	9/13 pSB1C3 EcoRI & SpeI	4.7 µL	9/15 Pcon-spinach-DT EcoRI & SpeI	4.7 µL	xx µL
experiment	9/13 pSB1C3 EcoRI & SpeI	4.7 µL	9/15 Pcon-apt12_1R-DT EcoRI & SpeI	2.0 µL	xx µL
experiment	9/13 pSB1C3 EcoRI & SpeI	4.7 µL	9/15 Pcon-pT181 attenuator-DT EcoRI & SpeI	3.0 µL	xx µL
experiment	9/13 pSB1C3 XbaI & PstI	1.9 µL	9/15 Pcon-pT181 anticodone-spinach-DT XbaI & PstI	4.4 µL	xx µL
experiment	9/13 pSB1C3 EcoRI & SpeI	4.7 µL	9/3 pT181 attenuator EcoRI & SpeI	3.8 µL	xx µL
experiment	9/6 Pcon SpeI & PstI	1.4 µL	9/15 RBS-lacZα-DT XbaI & PstI	2.0 µL	xx µL
experiment	9/14 pSB 4k5 EcoRI & SpeI	µL	9/15 Pcon-spinach-DT EcoRI & SpeI	9.4 µL	xx µL
experiment	9/15 Peet-RBS-lacZα-DT EcoRI & XbaI	2.6 µL	Pcon-RBS-tetR-DT EcoRI & SpeI	1.5 µL	xx µL

Restriction Enzyme Digestion

No Name

	DNA	XbaI	PstI	Buffer	BSA	MilliQ	total
9/13 pT181 attenuator XbaI & PstI	6.3 µL	1 µL	1 µL	3 µL	3 µL	15.7 µL	30 µL
NC	0.3 µL	0 µL	0 µL	1 µL	1 µL	7.7 µL	10 µL

	DNA	EcoRI	SpeI	Buffer	BSA	MilliQ	total
9/10 pSB1C3 EcoRI & SpeI	8.1 µL	1 µL	1 µL	3 µL	3 µL	13.9 µL	30 µL
NC	0.4 µL	0 µL	0 µL	1 µL	1 µL	7.6 µL	10 µL

	DNA	XbaI	PstI	Buffer	BSA	MilliQ	total
9/14 pSB4k5 XbaI & PstI	9.5 µL	1 µL	1 µL	3 µL	3 µL	12.5 µL	30 µL
NC	0.5 µL	0 µL	0 µL	1 µL	1 µL	7.5 µL	10 µL

	DNA	SpeI	PstI	Buffer	BSA	MilliQ	total
9/11 DT SpeI & PstI	10.4 µL	1 µL	1 µL	3 µL	3 µL	11.6 µL	30 µL
NC	0.5 µL	0 µL	0 µL	1 µL	1 µL	7.5 µL	10 µL

	DNA	SpeI	PstI	Buffer	BSA	MilliQ	total
9/6 apt12_1R-DT SpeI & PstI	7.9 µL	1 µL	1 µL	3 µL	3 µL	14.1 µL	30 µL
NC	0.4 µL	0 µL	0 µL	1 µL	1 µL	7.6 µL	10 µL

	DNA	SpeI	PstI	Buffer	BSA	MilliQ	total
9/10 spinach-DT SpeI & PstI	9.9 µL	1 µL	1 µL	3 µL	3 µL	12.1 µL	30 µL
NC	0.5 µL	0 µL	0 µL	1 µL	1 µL	7.5 µL	10 µL

Liquid Culture

No name

Sample	medium
pSB1C3
DT
Spinach-DT

Template:Kyoto/Notebook/Sep 15

From 2013.igem.org

Contents

Sep 15

Miniprep

Liquid Culture

Ligation

Restriction Enzyme Digestion

Electrophoresis

Gel Extraction

Ligation

Restriction Enzyme Digestion

Liquid Culture