Template:Kyoto/Notebook/Sep 16

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Contents

1 Sep 16

Sep 16

Liquid Culture

No name

Sample	medium
pSB1C3+P-KaiBC①	Plusgrow medium(+CP)
pSB1C3+P-KaiBC②	Plusgrow medium(+CP)

Colony PCR

No name

Sample	base pair
pSB1C3+P-KaiBC①	283
pSB1C3+P-KaiBC②	283

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
2min	30s	30s	18s	30cycles

Gel Extraction

じっけんしゃ

Lane	DNA	Enzyme
1	1kb ladder
2	pT181 attenuator	X+P
3	pT181 attenuator	X+P
4	pT181 attenuator	X+P
6	pSB1C3	E+S
7	pSB1C3	E+S
8	pSB1C3	E+S
10	pSB4K5	X+P
11	pSB4K5	X+P
12	pSB4K5	X+P

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

Name	concentration[µg/mL]	260/280	260/230
pT181 attenuator(X+P)	8.2	1.62	0.11
pSB1C3(E+S)	49.5	1.48	0.18
pSB4K5(X+P)	18.1	1.73	0.57
DT(S+P)	70.4	1.36	1.02
aptamer12-1R-DT(S+P)	42.0	1.82	0.47
spinach-DT(S+P)	36.8	1.79	0.61

Miniprep

No name

DNA	concentration[µg/mL]	260/280	260/230
Dx-lux1	136.7	1.46	1.28
RBS-laczα-DT	237.6	1.88	1.70
pSB4K5	148.4	1.55	1.09
Pcon-KBS-tetR-DT	126.6	1.73	1.36
9/11Plac1	160.0	1.78	1.28
9/15Plac2	144.0	1.73	1.81
9/14Plac	332.5	1.51	2.33
Ptet-RBS-laczα-DT	231.3	1.84	1.57

PCR

No name

DNA	9/6pT181 anti psB1C3(1)	KOD plus	10x buffer	dNTP	MgSO4	GG1_1C3_fwd primer	GG2_1C3_rev primer	MilliQ	total
400pg	1	0.5	2.5	2.5	1.5	0.75	0.75	15.5	25
800pg	2	0.5	2.5	2.5	1.5	0.75	0.75	14.5	25
1000pg	2.5	0.5	2.5	2.5	1.5	0.75	0.75	14	25
nega	0	0.5	2.5	2.5	1.5	0.75	0.75	16.5	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2min	10s	30s	30s	30cycles

DNA	9/6 12-1R aptamer-DT	KOD plus	10x buffer	dNTP	MgSO4	GG1_1C3_fwd primer	GG1_1C3_rev primer	MilliQ	total
400pg	1	0.5	2.5	2.5	1.5	0.75	0.75	15.5	25
800pg	2	0.5	2.5	2.5	1.5	0.75	0.75	14.5	25
1000pg	2.5	0.5	2.5	2.5	1.5	0.75	0.75	14	25
nega	0	0.5	2.5	2.5	1.5	0.75	0.75	16.5	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2min	10s	30s	30s	30cycles

Electrophoresis

じっけんしゃ

Lane	Sample
1	100bp ladder
2	anti 1000pg
3	anti 800pg
4	anti 400pg
5	tet aptamer-DT 1000pg
6	tet aptamer-DT 800pg
7	tet aptamer-DT 400pg
8	nega

File:Igku xxxxxx.xxx

Restriction Enzyme Digestion

じっけんしゃ

	9/16 Pcon-RBS-tetR-DT(µL)	E(µL)	S(µL)	X(µL)	P(µL)	Buffer(µL)	BSA(µL)	MilliQ(µL)	total(µL)
2 cuts	15.8	0	0	1	1	3	3	6.2	30
NC	0.8	0	0	0	0	1	1	7.2	10

	9/16 pSB4K5(µL)	E(µL)	S(µL)	X(µL)	P(µL)	Buffer(µL)	BSA(µL)	MilliQ(µL)	total(µL)
2 cuts	13.5	0	0	1	1	3	3	8.5	30
NC	0.7	0	0	0	0	1	1	7.3	10

	8/22 pSB1C3(µL)	E(µL)	S(µL)	X(µL)	P(µL)	Buffer(µL)	BSA(µL)	MilliQ(µL)	total(µL)
2 cuts	18.5	1	0	0	1	3	3	3.5	30
NC	0.9	0	0	0	0	1	1	7.1	10

	8/22 pSB1C3(µL)	E(µL)	S(µL)	X(µL)	P(µL)	Buffer(µL)	BSA(µL)	MilliQ(µL)	total(µL)
2 cuts	18.5	1	1	0	0	3	3	3.5	30
NC	0.9	0	0	0	0	1	1	7.1	10

	8/21 pT181 attenuator2(µL)	E(µL)	S(µL)	X(µL)	P(µL)	Buffer(µL)	BSA(µL)	MilliQ(µL)	total(µL)
2 cuts	7.4	1	1	0	0	3	3	14.6	30
NC	0.4	0	0	0	0	1	1	7.6	10

Ligation

じっけんしゃ

state	Vector		Inserter		Ligation High ver.2
experiment	Pcon-pT181anti-spin-DT(X+P)	5.0µg/mL	pSB4K5(X+P)	18.1µg/mL	xx µg/mL
experiment	pC181 attenuator(X+P)	8.2µg/mL	pSB1C3(X+P)	26.0µg/mL	xx µg/mL

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