Template:Kyoto/Notebook/Aug 22

From 2013.igem.org

(Difference between revisions)

Revision as of 02:45, 25 September 2013

Aug 22

Liquid culture

Nakamoto

Sample	Medium
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (1)	8/21 Plusgrow medium (+Amp)
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (2)	8/21 Plusgrow medium (+Amp)
8/21 Pcon-RBS-GFP-DT control (1)	8/21 Plusgrow medium (+Amp)
8/21 Pcon-RBS-GFP-DT control (2)	8/21 Plusgrow medium (+Amp)
8/21 RBS-lysis1 (1)	8/21 Plusgrow medium (+Amp)
8/21 RBS-lysis1 (2)	8/21 Plusgrow medium (+Amp)
8/21 RBS control (1)	8/21 Plusgrow medium (+Amp)
8/21 RBS control (2)	8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm)-RBS-tetR-DT (1)	8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm)-RBS-tetR-DT (2)	8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm) control (1)	8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm) control (2)	8/21 Plusgrow medium (+Amp)
8/21 Plux-RBS-GFP-DT (1)	8/21 Plusgrow medium (+Amp)
8/21 pSB1C3(BBa_J04450) (1)	8/21 Plusgrow medium (+Amp)
8/21 pSB1C3(BBa_J04450) (2)	8/21 Plusgrow medium (+Amp)

incubate 37°C 10hour

Colony PCR

Kojima

Sample	base pair
8/21 RBS-lysis1(1)	400
8/21 RBS-lysis1(2)	400
8/21 RBS control(1)	--
8/21 RBS control(2)	--
negative control	--

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

Sample	base pair
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (1)	2143
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (2)	2143
8/21 Pcon-RBS-luxR control(1)	--
8/21 Pcon-RBS-luxR control(2)	--
negative control	--

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5min	30s	30s	2min	30cycles

Sample	base pair
8/21 Ptet-RBS-tetR-DT (1)	1216
8/21 Ptet-RBS-tetR-DT (2)	1216
8/21 Ptet control (1)	--
8/21 Ptet control (2)	--
8/21 Plux-RBS-GFP-DT (1)	1227
8/21 pSB1C3(BBa_J04450) (1)	1353
8/21 pSB1C3(BBa_J04450) (2)	1353
negative control	--

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5min	30s	30s	1min	30cycles

Miniprep

Ashida

DNA	concentration[µg/mL]	260/280	260/230
8/22 RBS-lysis (1)	128	1.77	2.29
8/22 RBS-lysis (2)	96	1.80	2.07
8/22 pSB1C3 (1)	108	1.55	1.45
8/22 pSB1C3 (2)	130	1.69	1.76

Electrophoresis

No name

Lane	Sample	Enzyme1	Enzyme2
1	100bp	--	--
2	8/22 pT181 attenuator (1)	EcoRI	SpeI
3	8/22 pT181 attenuator (1)	EcoRI	--
4	8/22 pT181 attenuator (1)	--	SpeI
5	8/22 pT181 attenuator (1)	--	--
6	8/22 pT181 attenuator (1)	XbaI	PstI
7	8/22 pT181 attenuator (1)	XbaI	--
8	8/22 pT181 attenuator (1)	--	PstI
9	8/22 pT181 attenuator (1)	--	--
10	8/22 Fusion6 antisense (1)	EcoRI	SpeI
11	8/22 Fusion6 antisense (1)	EcoRI	--
12	8/22 Fusion6 antisense (1)	--	SpeI
13	8/22 Fusion6 antisense (1)	--	--
14	100bp	--	--

Lane	Sample	Enzyme1	Enzyme2
1	100bp	--	--
2	8/22 Fusion6 antisense (1)	XbaI	PstI
3	8/22 Fusion6 antisense (1)	XbaI	--
4	8/22 Fusion6 antisense (1)	--	PstI
5	8/22 Fusion6 antisense (1)	--	--
6	8/22 Fusion1 antisense (1)	EcoRI	SpeI
7	8/22 Fusion1 antisense (1)	EcoRI	--
8	8/22 Fusion1 antisense (1)	--	SpeI
9	8/22 Fusion1 antisense (1)	--	--
10	8/22 Fusion1 antisense (1)	XbaI	PstI
11	8/22 Fusion1 antisense (1)	XbaI	--
12	8/22 Fusion1 antisense (1)	--	PstI
13	8/22 Fusion1 antisense (1)	--	--
14	100bp	--	--

@@ Line 110: / Line 110: @@
 |5min||30s||30s||1min||30cycles
 |}
-</div>
-===Restriction Enzyme Digestion===
-<div class="experiment">
-<span class="author">Tatsui, Kojima, Honda and Ashida</span>
-{| class="wikitable"
-! ||Fusion1 antisense (1) (300 &micro;g/mL)||EcoRI||SpeI||buffer H||MilliQ||total
-|-
-|2 cuts||3.3||1||1||3||21.7||30
-|-
-|1 cut||0.7||0.2||0||1||8.1||10
-|-
-|1 cut||0.7||0||0.2||1||8.1||10
-|-
-|NC||0.7||0||0||1||8.3||10
-|}
-{| class="wikitable"
-! ||Fusion1 antisense (1) (300 &micro;g/mL)||XbaI||PstI||buffer M||BSA||MilliQ||total
-|-
-|2 cuts||3.3||1||1||3||3||18.7||30
-|-
-|1 cut||0.7||0.2||0||1||1||7.1||10
-|-
-|1 cut||0.7||0||0.2||1||1||7.1||10
-|-
-|NC||0.7||0||0||1||1||7.3||10
-|}
-{| class="wikitable"
-! ||Fusion6 antisense (1) (336 &micro;g/mL)||EcoRI||SpeI||buffer H||MilliQ||total
-|-
-|2 cuts||3||1||1||3||22||30
-|-
-|1 cut||0.6||0.2||0||1||8.2||10
-|-
-|1 cut||0.6||0||0.2||1||8.2||10
-|-
-|NC||0.6||0||0||1||8.4||10
-|}
-{| class="wikitable"
-! ||Fusion6 antisense (1) (336 &micro;g/mL)||XbaI||PstI||buffer M||BSA||MilliQ||total
-|-
-|2 cuts||3||1||1||3||3||19||30
-|-
-|1 cut||0.6||0.2||0||1||1||7.2||10
-|-
-|1 cut||0.6||0||0.2||1||1||7.2||10
-|-
-|NC||0.6||0||0||1||1||7.4||10
-|}
-{| class="wikitable"
-! ||pT181 antisense (1) (178 &micro;g/mL)||EcoRI||SpeI||buffer H||MilliQ||total
-|-
-|2 cuts||5.6||1||1||3||19.4||30
-|-
-|1 cut||1.1||0.2||0||1||7.7||10
-|-
-|1 cut||1.1||0||0.2||1||7.7||10
-|-
-|NC||1.1||0||0||1||7.9||10
-|}
-{| class="wikitable"
-! ||pT181 antisense (1) (178 &micro;g/mL)||XbaI||PstI||buffer M||BSA||MilliQ||total
-|-
-|2 cuts||5.6||1||1||3||3||16.4||30
-|-
-|1 cut||1.1||0.2||0||1||1||6.7||10
-|-
-|1 cut||1.1||0||0.2||1||1||6.7||10
-|-
-|NC||1.1||0||0||1||1||6.9||10
-|}
-{| class="wikitable"
-! ||pT181 attenuator (1)||EcoRI||SpeI||buffer H||MilliQ||total
-|-
-|2 cuts||3||1||1||3||22||30
-|-
-|1 cut||0.6||0.2||0||1||8.2||10
-|-
-|1 cut||0.6||0||0.2||1||8.2||10
-|-
-|NC||0.6||0||0||1||8.4||10
-|}
-{| class="wikitable"
-! ||pT181 attenuator (1)||XbaI||PstI||buffer M||BSA||MilliQ||total
-|-
-|2 cuts||3||1||1||3||3||19||30
-|-
-|1 cut||0.6||0.2||0||1||1||7.2||10
-|-
-|1 cut||0.6||0||0.2||1||1||7.2||10
-|-
-|NC||0.6||0||0||1||1||7.4||10
-|}
-{| class="wikitable"
-! ||Fusion1 attenuator (1)||EcoRI||SpeI||buffer H||MilliQ||total
-|-
-|2 cuts||3.3||1||1||3||21.7||30
-|-
-|1 cut||0.7||0.2||0||1||8.1||10
-|-
-|1 cut||0.7||0||0.2||1||8.1||10
-|-
-|NC||0.7||0||0||1||8.3||10
-|}
-{| class="wikitable"
-! ||Fusion1 attenuator (1)||XbaI||PstI||buffer M||BSA||MilliQ||total
-|-
-|2 cuts||3.3||1||1||3||3||18.7||30
-|-
-|1 cut||0.7||0.2||0||1||1||7.1||10
-|-
-|1 cut||0.7||0||0.2||1||1||7.1||10
-|-
-|NC||0.7||0||0||1||1||7.3||10
-|}
-{| class="wikitable"
-! ||Fusion3m2 attenuator (1)||EcoRI||SpeI||buffer H||MilliQ||total
-|-
-|2 cuts||3||1||1||3||22||30
-|-
-|1 cut||0.6||0.2||0||1||8.2||10
-|-
-|1 cut||0.6||0||0.2||1||8.2||10
-|-
-|NC||0.6||0||0||1||8.4||10
-|}
-{| class="wikitable"
-! ||Fusion3m2 attenuator (1)||XbaI||PstI||buffer M||BSA||MilliQ||total
-|-
-|2 cuts||3||1||1||3||3||19||30
-|-
-|1 cut||0.6||0.2||0||1||1||7.2||10
-|-
-|1 cut||0.6||0||0.2||1||1||7.2||10
-|-
-|NC||0.6||0||0||1||1||7.4||10
-|}
-{| class="wikitable"
-! ||Spinach (2)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
-|-
-|2 cuts||2.3||0.5||0.5||3||0.3||23.4||30
-|-
-|1 cut||0.5||0.1||0||1||0.1||8.3||10
-|-
-|1 cut||0.5||0||0.1||1||0.1||8.3||10
-|-
-|NC||0.5||0||0||1||0.1||8.4||10
-|}
-{| class="wikitable"
-! ||tetR aptamer 12_P (2)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
-|-
-|2 cuts||2.7||0.5||0.5||3||0.3||23||30
-|-
-|1 cut||0.5||0.1||0||1||0.1||8.3||10
-|-
-|1 cut||0.5||0||0.1||1||0.1||8.3||10
-|-
-|NC||0.5||0||0||1||0.1||8.4||10
-|}
-{| class="wikitable"
-! ||tetR aptamer 12_1R (2)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
-|-
-|2 cuts||2.6||0.5||0.5||3||0.3||23.1||30
-|-
-|1 cut||0.5||0.1||0||1||0.1||8.3||10
-|-
-|1 cut||0.5||0||0.1||1||0.1||8.3||10
-|-
-|NC||0.5||0||0||1||0.1||8.4||10
-|}
-{| class="wikitable"
-! ||tetR aptamer 12_1M (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
-|-
-|2 cuts||2.7||0.5||0.5||3||0.3||23||30
-|-
-|1 cut||0.5||0.1||0||1||0.1||8.3||10
-|-
-|1 cut||0.5||0||0.1||1||0.1||8.3||10
-|-
-|NC||0.5||0||0||1||0.1||8.4||10
-|}
-{| class="wikitable"
-! ||J23100||SpeI||PstI||buffer B||BSA||MilliQ||total
-|-
-|2 cuts||7||0.5||0.5||3||0.3||18.7||30
-|-
-|1 cut||1.4||0.1||0||1||0.1||7.4||10
-|-
-|1 cut||1.4||0||0.1||1||0.1||7.4||10
-|-
-|NC||1.4||0||0||1||0.1||7.5||10
-|}
-{| class="wikitable"
-! ||Plac (2)||SpeI||PstI||buffer B||BSA||MilliQ||total
-|-
-|2 cuts||11.2||0.5||0.5||3||0.3||14.5||30
-|-
-|1 cut||2.2||0.1||0||1||0.1||6.6||10
-|-
-|1 cut||2.2||0||0.1||1||0.1||6.6||10
-|-
-|NC||2.2||0||0||1||0.1||6.7||10
-|}
 </div>

Template:Kyoto/Notebook/Aug 22

From 2013.igem.org

Revision as of 02:45, 25 September 2013

Contents

Aug 22

Liquid culture

Colony PCR

Miniprep

Electrophoresis