Template:Kyoto/Notebook/Sep 9

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Contents

1 Sep 9

Sep 9

PCR

Nakagawa

KaiA(plasmid)	10x buffer	dNTPs	MgSO4	Primer(661-RBS-KaiA-fwd)	Primer(kaiA-662-rev-ver2)	Kod-plus ver2	MilliQ	total
1.0	2.5	2.5	1.5	0.75	0.75	0.5	15.5	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	58.5°C	68°C	--
2min	10s	30s	30s	30cycles

KaiB(plasmid)	10x buffer	dNTPs	MgSO4	Primer(661-RBS-KaiA-fwd-ver2)	Primer(kaiB-663-rev-ver2)	Kod-plus ver2	MilliQ	total
1.0	2.5	2.5	1.5	0.75	0.75	0.5	15.5	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	58.5°C	68°C	--
2min	10s	30s	30s	30cycles

P-KaiBC(PCR products)	10x buffer	dNTPs	MgSO4	Primer(prefix-P-kaiBC-fwd-ver2)	Primer(P-KaiBC-suffix-rev-verw)	Kod-plus ver2	MilliQ	total
0.7	2.5	2.5	1.5	0.75	0.75	0.5	15.8	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	58.5°C	68°C	--
2min	10s	30s	30s	30cycles

DT(plasmid)	10x buffer	dNTPs	MgSO4	Primer(664-DT-fwd-ver2)	Primer(DT-suffix-660-rev-ver2)	Kod-plus ver2	MilliQ	total
1.0	2.5	2.5	1.5	0.75	0.75	0.5	15.5	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	58.5°C	68°C	--
2min	10s	30s	30s	30cycles

Miniprep

No name

DNA	concentration[µg/mL]	260/280	260/230
9/8 pSB1C3	83.2	1.45	0.76
9/8 RBS-lysis2-DT	72.8	1.67	0.97
9/8 spinach-DT	152.6	1.88	1.97
Nuclease-Free water	-0.4	-14.30	0.25

Restriction Enzyme Digestion

じっけんしゃ

	DNA	restricion enzyme	restriction enzyme	Buffer	BSA	MilliQ	total
2 cuts	（°°） µL	µL	µL	µL	µL	µL	µL
NC	µL	µL	µL	µL	µL	µL	µL

Transformation

じっけんしゃ

Name	Sample	Competent Cells	Total	Plate
いでんし	なんちゃらµL	ごにょごにょµL	ほにゃららµL	こーせーぶっしつ
いかどうよう

Ligation

No Name

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

Colony PCR

No name

Sample	base pair
9/8 Plac-spinach-DT-(1)	659

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5m	30s	30s	40s	30cycles

Sample	base pair
9/8 Plac-spinach-DT-(2)	659

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5m	30s	30s	40s	30cycles

Sample	base pair
9/8 Pcon-pt181atte-DT-(1)	584

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5m	30s	30s	40s	30cycles

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