Template:Kyoto/Notebook/Sep 8

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Sep 8

Gel Extraction

No name

Lane	DNA	Enzyme
1	100bp ladder
2	RBS-lysis1-DT	XbaI & PstI
3	RBS-lysis1-DT	XbaI & PstI
5	RBS-lysis3-DT	XbaI & PstI
6	RBS-lysis3-DT	XbaI & PstI

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

Name	concentration[µg/mL]	260/280	260/230
RBS-lysis1-DT (XbaI & PstI)	5.3	1.98	0.06
RBS-lysis3-DT (XbaI & PstI)	5.3	1.98	0.06

Lane	DNA	Enzyme
1	100bp ladder
3	Pcon-pT181 attenuator	EcoRI & SpeI
4	Pcon-pT181 attenuator	EcoRI & SpeI
6	Spinach-DT	XbaI & PstI
7	Spinach-DT	XbaI & PstI
9	RBS-lacZα-DT	XbaI & PstI
10	RBS-lacZα-DT	XbaI & PstI

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

Name	concentration[µg/mL]	260/280	260/230
Pcon-pT181 attenuator (EcoRI & SpeI)	5.2	1.82	0.36
Spinach-DT (XbaI & PstI)	8.0	1.87	0.25
RBS-lacZα-DT (XbaI & PstI)	10.3	1.71	0.54

Lane	DNA	Enzyme
1	1kbp ladder
2	Ptet-pT181 antisense	EcoRI & PstI
3	Ptet-pT181 antisense	EcoRI & PstI
5	pSB4K5	EcoRI & SpeI
6	pSB4K5	EcoRI & SpeI

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

Name	concentration[µg/mL]	260/280	260/230
Ptet-pT181 antisense(EcoRI & PstI)	30.6	1.84	1.16
pSB4K5 (EcoRI & SpeI)	21.8	1.87	0.98

Lane	DNA	Enzyme
1	1kbp ladder
2	Pcon-pT181 attenuator	SpeI & PstI
3	Pcon-pT181 attenuator	SpeI & PstI
5	Plac	SpeI & PstI
6	Plac	SpeI & PstI
8	pSB1C3	EcoRI & SpeI
9	pSB1C3	EcoRI & SpeI
11	pSB1C3	XbaI & PstI
12	pSB1C3	XbaI & PstI

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

Name	concentration[µg/mL]	260/280	260/230
Pcon-pT181 attenuator (SpeI & PstI)	14.5	1.91	1.09
Plac (SpeI & PstI)	11.6	1.74	1.19
pSB1C3 (EcoRI & SpeI)	5.7	1.74	0.34
pSB1C3 (XbaI & PstI)	5.6	2.00	0.37

Miniprep

No name

DNA	concentration[µg/mL]	260/280	260/230
Pcon-pT181 anti	243.0	1.90	1.81
Pcon-Spinach-DT	253.7	1.57	1.51

Restriction Enzyme Digestion

じっけんしゃ

	9/8 Pcon-pT181 anti	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	8.2µL	0µL	1µL	0µL	1µL	3µL	3µL	13.8µL	30µL
NC	0.4µL	0µL	0µL	0µL	0µL	1µL	1µL	7.6µL	10µL

	9/8 Pcon-spin-DT	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	7.9µL	1µL	1µL	0µL	0µL	3µL	3µL	14.1µL	30µL
NC	0.4µL	0µL	0µL	0µL	0µL	1µL	1µL	7.6µL	10µL

	8/21 F1 atte	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	6.5µL	0µL	0µL	1µL	1µL	3µL	3µL	15.5µL	30µL
NC	0.3µL	0µL	0µL	0µL	0µL	1µL	1µL	7.7µL	10µL

	8/21 F3m2 atte	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	6.0µL	0µL	0µL	1µL	1µL	3µL	3µL	16µL	30µL
NC	0.3µL	0µL	0µL	0µL	0µL	1µL	1µL	7.7µL	10µL

	8/21 F1 anti	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	6.7µL	0µL	0µL	1µL	1µL	3µL	3µL	15.3µL	30µL
NC	0.3µL	0µL	0µL	0µL	0µL	1µL	1µL	7.7µL	10µL

	8/21 F6 anti	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	6.0µL	0µL	0µL	1µL	1µL	3µL	3µL	16µL	30µL
NC	0.3µL	0µL	0µL	0µL	0µL	1µL	1µL	7.7µL	10µL

	8/21 tetR aptamer 12_1M	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	5.3µL	1µL	1µL	0µL	0µL	3µL	3µL	16.7µL	30µL
NC	0.3µL	0µL	0µL	0µL	0µL	1µL	1µL	7.7µL	10µL

	8/21 tetR aptamer 12_P	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	13.3µL	1µL	1µL	0µL	0µL	3µL	3µL	8.7µL	30µL
NC	0.7µL	0µL	0µL	0µL	0µL	1µL	1µL	7.3µL	10µL

	9/4 spin-DT	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	5.4µL	0µL	0µL	1µL	1µL	3µL	3µL	16.6µL	30µL
NC	0.3µL	0µL	0µL	0µL	0µL	1µL	1µL	7.7µL	10µL

	9/7 RBS-lysis1-DT	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	10µL	0µL	0µL	1µL	1µL	3µL	3µL	12µL	30µL
NC	0.5µL	0µL	0µL	0µL	0µL	1µL	1µL	7.5µL	10µL

	9/6 RBS-lysis2-DT	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	13µL	0µL	0µL	1µL	1µL	3µL	3µL	9µL	30µL
NC	0.7µL	0µL	0µL	0µL	0µL	1µL	1µL	7.3µL	10µL

	9/7 RBS-lysis3-DT	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	10µL	0µL	0µL	1µL	1µL	3µL	3µL	12µL	30µL
NC	0.5µL	0µL	0µL	0µL	0µL	1µL	1µL	7.5µL	10µL

	8/20 J23100 RBS-lacZα-DT-(1)	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	7µL	0.5µL	0.5µL	0µL	0µL	3µL	3µL	16µL	30µL
NC	0.5µL	0µL	0µL	0µL	0µL	1µL	1µL	7.5µL	10µL

	8/20 RBS-lacZα-DT	E	S	X	P	BSA	Buffer	MilliQ	total
2 cuts	10µL	0.5µL	0.5µL	0µL	0µL	3µL	3µL	13µL	30µL
NC	0.5µL	0µL	0µL	0µL	0µL	1µL	1µL	7.5µL	10µL

Transformation

じっけんしゃ

Name	Sample(µL)	Competent Cells(µL)	Total(µL)	Plate
9/8 Pcont+aptamer 12-1R-DT	1	10	11	Amp
9/8 Pcon+RBS-lacZα-DT	1	10	11	Amp
9/8 Pcon-pT181 attenuater+aptamer 12-1R-DT	1	10	11	Amp
9/8 Plac+RBS-lysis1-DT	2	20	22	CP
9/8 Plac+RBS-lysis2-DT	2	20	22	CP
9/8 Plac+aptamer 12-1R-DT	2	20	22	CP
9/8 Plac+spinach	2	20	22	CP
9/8 Plac+pT181 attenuater	2	20	22	CP
9/8 Plux+RBS-lysis1-DT	1	10	11	CP
9/8 Plux+RBS-lysis2-DT	1	10	11	CP
9/8 Ptet+RBS-lacZα-DT	1	10	11	CP
9/8 Ptet-pT181 anti+spinach-DT	1	10	11	CP
9/8 Pcon-pT181 attenuater+DT	1	10	11	CP

Electrophoresis

じっけんしゃ

Lane	Sample	Enzyme1	Enzyme2
（°Д°）

Lane	Sample	Enzyme1	Enzyme2
（°Д°）

Lane	Sample	Enzyme1	Enzyme2
（°Д°）

Gel Extraction

じっけんしゃ

Lane	DNA	Enzyme
（＞ω・）b

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

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

Lane	DNA	Enzyme
（＞ω・）b

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

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

Lane	DNA	Enzyme
（＞ω・）b

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

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

Lane	DNA	Enzyme
（＞ω・）b

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

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

Lane	DNA	Enzyme
（＞ω・）b

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

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

Ligation

じっけんしゃ

state	Vector		Inserter		Ligation High ver.2
experiment	name	xx µg/mL	name	xx µg/mL	xx µg/mL
NC	name	xx µg/mL	name	xx µg/mL	xx µg/mL

state	Vector		Inserter		Ligation High ver.2
experiment	name	xx µg/mL	name	xx µg/mL	xx µg/mL
NC	name	xx µg/mL	name	xx µg/mL	xx µg/mL

PCR

No name

genome DNA	KOD plus	10x buffer	dNTP	MgSO4	SasA_fwd primer	SasA_rev primer	MilliQ	total
	0.5	2.5	2.5	1.5	0.75	0.75		25

PreDenature	Denature	Annealing	Extension	cycle
°C	°C	°C	°C	--
（◎Д◎）

Gel Extraction

じっけんしゃ

Lane	DNA	Enzyme
（＞ω・）b

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

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

Lane	DNA	Enzyme
（＞ω・）b

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

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

Template:Kyoto/Notebook/Sep 8

From 2013.igem.org

Contents

Sep 8

Gel Extraction

Miniprep

Restriction Enzyme Digestion

Transformation

Electrophoresis

Gel Extraction

Ligation

PCR

Gel Extraction