Template:Kyoto/Notebook/Sep 17

From 2013.igem.org

(Difference between revisions)

Revision as of 03:12, 24 September 2013

Sep 17

Liquid Culture

Tatsui and Hirano

Sample	medium
Spi-DT	LB medium(+CP)
Pcon-Spi-DT,Pcon-pT181-anti-spi-DT	LB medium(+Amp)

PCR

Hirano

Pcon-tet ap-DT(VF)	BigDye	5x buffer	primer(VF)	MilliQ	total
1.3	0.5	1.75	0.5	0.45	10.5

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

Pcon-tet ap-DT(VR)	BigDye	5x buffer	primer(VR)	MilliQ	total
1.3	0.5	1.75	0.5	0.45	10.5

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

Ligation

No Name

state	Vector		Inserter		Ligation High ver.2
experiment	8/28 Plux(S+P)	45µg/mL	9/13 RBS-lysis1-DT (X+P)	7.8µg/mL	xx µg/mL
experiment	8/28 Plux(S+P)	45µg/mL	9/13 RBS-lysis2-DT (X+P)	11.8µg/mL	xx µg/mL
experiment	8/28 Plux(S+P)	45µg/mL	9/13 RBS-lysis3-DT (X+P)	11.8µg/mL	xx µg/mL
experiment	9/14 Plac(S+P)	21µg/mL	9/16 PT181 atte-DT (X+P)	8.2µg/mL	xx µg/mL
experiment	9/14 Plac(S+P)	21µg/mL	9/3 PT181 anti (X+P)	6.6µg/mL	xx µg/mL
experiment	9/14 Plac(S+P)	21µg/mL	9/10 apt12-1R-DT(X+P)	15.6µg/mL	xx µg/mL
experiment	9/13 pSB1C3(X+P)	26.0µg/mL	9/15 Pcon-PT181 anti-spin-DT(X+P)	5.0µg/mL	xx µg/mL
experiment	9/16 pSB1C3(E+S)	49.5µg/mL	9/15 Pcon-spin-DT(E+S)	9.0µg/mL	xx µg/mL
experiment	9/16 pSB1C3(E+S)	49.5µg/mL	9/15 Pcon-RBS-tetR-DT(E+S)	59.0µg/mL	xx µg/mL
experiment	9/16 pSB1C3(E+S)	49.5µg/mL	9/15 Pcon-PT181-atte-DT(E+S)	21.0µg/mL	xx µg/mL
experiment	9/16 pSB1C3(E+S)	49.5µg/mL	9/15 Pcon-apt12-112-DT(E+S)	17.0µg/mL	xx µg/mL

Gel Extraction

No Name

Lane	DNA	Enzyme
1	1kbpladder	--
2	Pt181 atte -(2)	E+S
3
4

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

Name	concentration[µg/mL]	260/280	260/230
PT181-atte E+S	9.2	1.15	0.12

Lane	DNA	Enzyme
1	1kbpladder	--
2	pSB1C3	E+S
3
4

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

Name	concentration[µg/mL]	260/280	260/230
pSB1C3	10.0	1.84	0.40

Lane	DNA	Enzyme
1	1kbpladder	--
2	pSB4K5	X+P
3
4

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

Name	concentration[µg/mL]	260/280	260/230
pSB4K5	7.6	1.81	0.32

Lane	DNA	Enzyme
1	1kbpladder	--
2	Pcon-RBS-tetR-DT	X+P
3
4

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

Name	concentration[µg/mL]	260/280	260/230
Pcon-RBS-tetR-DT	8.0	2.40	1.14

Lane	DNA	Enzyme
1	1kbpladder	--
2	pSB1C3	E+P
3
4

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

Name	concentration[µg/mL]	260/280	260/230
pSB1C3	9.1	2.17	0.35

PCR

Kinose

pC KaiABC(100pg)	Prime STAR(2x)	Primer(KaiA-Bsai MuT-Fwd)	Primer(KaiA-Bsai MuT-Rev)	MilliQ	total
1.0	25.0	1.0	1.0	22.0	50.0

PreDenature	Denature	Annealing	Extension	cycle
96°C	98°C	65°C	72°C	--
5m	10s	15s	40s	30cycles

pC KaiABC(150pg)	Prime STAR(2x)	Primer(KaiA-Bsai MuT-Fwd)	Primer(KaiA-Bsai MuT-Rev)	MilliQ	total
1.5	25.0	1.0	1.0	21.5	50.0

PreDenature	Denature	Annealing	Extension	cycle
96°C	98°C	65°C	72°C	--
5m	10s	15s	40s	30cycles

</div>

pC KaiABC(200pg)	Prime STAR(2x)	Primer(KaiA-Bsai MuT-Fwd)	Primer(KaiA-Bsai MuT-Rev)	MilliQ	total
2	25.0	1.0	1.0	21.0	50.0

PreDenature	Denature	Annealing	Extension	cycle
96°C	98°C	65°C	72°C	--
5m	10s	15s	40s	30cycles

@@ Line 69: / Line 69: @@
 |-
 |experiment||9/16 pSB1C3(E+S)||49.5&micro;g/mL||9/15 Pcon-RBS-tetR-DT(E+S)||59.0&micro;g/mL||xx &micro;g/mL
+|-
+|experiment||9/16 pSB1C3(E+S)||49.5&micro;g/mL||9/15 Pcon-PT181-atte-DT(E+S)||21.0&micro;g/mL||xx &micro;g/mL
+|-
+|experiment||9/16 pSB1C3(E+S)||49.5&micro;g/mL||9/15 Pcon-apt12-112-DT(E+S)||17.0&micro;g/mL||xx &micro;g/mL
 |}
 </div>

Template:Kyoto/Notebook/Sep 17

From 2013.igem.org

Revision as of 03:12, 24 September 2013

Contents

Sep 17

Liquid Culture

PCR

Ligation

Gel Extraction

PCR