Template:Kyoto/Notebook/Sep 3

From 2013.igem.org

(Difference between revisions)

Revision as of 10:37, 25 September 2013

Sep 3

Electrophoresis

Kozima

Lane	Sample
1	100bp ladder
2	pT181 attenuater(2)-DT2(738)
3	Ptet-RBS-lacZα-DT-2(756)
4	Ptet-RBS-lacZα-DT-3
5	Ptet-RBS-lacZα-DT-4
6	Ptet-RBS-lacZα-DT-5
7	Ptet-RBS-lacZα-DT-6
8	RBS-lysis2-DT-2(985)
9	RBS-lysis2-DT-3
10	100bp ladder

Miniprep

Tatsui

DNA	concentration[µg/mL]	260/280	260/230
9/2 14:00 pT181 attenuator	418.4	1.82	1.80
9/2 14:00 pT181 antisense	362.2	1.92	1.96
9/2 14:00 Spinach	438.3	1.86	1.99
9/2 14:00 tetR-apt 12_1R	567.5	1.90	1.99
9/2 21:00 pT181 attenuater	320.9	1.90	2.15
9/2 21:00 Spinach	457.0	1.88	2.24

Liquid Culture

Kozima

Sample	medium
Spinach-DT(9/2 master plate)	plusgrow(CP)4ml

Restriction Enzyme Digestion

Kozima

	DNA	EcoR1	Spe1	Xba1	Pst1	10xBuffer B(EcoR1&Spe1)	10xBuffer D(Xba1&Pst1)	100xBSA	MilliQ	total
9/3 pT181attenuater 14:00 418.4ng/µL	4.8µL	1.0µL	1.0µL	0µL	0µL	3µL	0µL	0.3µL	19.9µL	30µL
9/3 pT181attenuater 14:00 418.4ng/µL	0.2µL	0µL	0µL	0µL	0µL	1µL	0µL	0.1µL	8.7µL	10µL
9/3 pT181attenuater 14:00 418.4ng/µL	4.8µL	0µL	0µL	1.0µL	1.0µL	0µL	3µL	0.3µL	19.9µL	30µL
9/3 pT181attenuater 14:00 418.4ng/µL	0.2µL	0µL	0µL	0µL	0µL	0µL	1µL	0.1µL	8.7µL	10µL

	DNA	EcoR1	Spe1	Xba1	Pst1	10xBuffer B(EcoR1&Spe1)	10xBuffer D(Xba1&Pst1)	100xBSA	MilliQ	total
9/3 pT181antisense 14:00 362.2ng/µL	5.5µL	1.0µL	1.0µL	0µL	0µL	3µL	0µL	0.3µL	19.2µL	30µL
9/3 pT181attenuater 14:00 418.4ng/µL	0.3µL	0µL	0µL	0µL	0µL	1µL	0µL	0.1µL	8.6µL	10µL
9/3 pT181attenuater 14:00 418.4ng/µL	5.5µL	0µL	0µL	1.0µL	1.0µL	0µL	3µL	0.3µL	19.2µL	30µL
9/3 pT181attenuater 14:00 418.4ng/µL	0.3µL	0µL	0µL	0µL	0µL	0µL	1µL	0.1µL	8.6µL	10µL

	DNA	EcoR1	Spe1	10xBuffer B	100xBSA	MilliQ	total
9/3 apt 12_1R 14:00 507.5ng/µL	3.9µL	1.0µL	1.0µL	3µL	0.3µL	20.8µL	30µL
9/3 apt 12_1R 14:00 507.5ng/µL	0.2µL	0µL	0µL	1µL	0.1µL	8.7µL	10µL

	DNA	EcoR1	Xba1	10xBuffer D	100xBSA	MilliQ	total
8/29 DT 166.2ng/µL	12µL	1.0µL	1.0µL	3µL	0.3µL	12.7µL	30µL
8/29 DT 166.2ng/µL	0.6µL	0µL	0µL	1µL	0.1µL	8.3µL	10µL

	DNA	EcoR1	Spe1	10xBuffer B	100xBSA	MilliQ	total
8/22 RBS-lysis1(2) 96ng/µL	21µL	1.0µL	1.0µL	3µL	0.3µL	3.7µL	30µL
8/22 RBS-lysis1(2) 96ng/µL	1.0µL	0µL	0µL	1µL	0.1µL	7.9µL	10µL

	DNA	EcoR1	Spe1	10xBuffer B	100xBSA	MilliQ	total
8/30 RBS-lysis2 220ng/µL	9.1µL	1.0µL	1.0µL	3µL	0.3µL	15.6µL	30µL
8/30 RBS-lysis2 220ng/µL	0.5µL	0µL	0µL	1µL	0.1µL	8.4µL	10µL

	DNA	EcoR1	Spe1	10xBuffer B	100xBSA	MilliQ	total
8/20 RBS-lysis3(1) 282ng/µL	7.1µL	1.0µL	1.0µL	3µL	0.3µL	17.6µL	30µL
8/20 RBS-lysis3(1) 282ng/µL	0.4µL	0µL	0µL	1µL	0.1µL	8.5µL	10µL

@@ Line 69: / Line 69: @@
 ! ||DNA||EcoR1||Spe1||Xba1||Pst1||10xBuffer B(EcoR1&Spe1)||10xBuffer D(Xba1&Pst1)||100xBSA||MilliQ||total
 |-
-|pT181attenuater 14:00 418.4ng/&micro;L|| 4.8&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 0&micro;L|| 3&micro;L|| 0&micro;L|| 0.3&micro;L|| 19.9&micro;L|| 30&micro;L
+|9/3 pT181attenuater 14:00 418.4ng/&micro;L|| 4.8&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 0&micro;L|| 3&micro;L|| 0&micro;L|| 0.3&micro;L|| 19.9&micro;L|| 30&micro;L
 |-
-|pT181attenuater 14:00 418.4ng/&micro;L|| 0.2&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0&micro;L|| 0.1&micro;L|| 8.7&micro;L|| 10&micro;L
+|9/3 pT181attenuater 14:00 418.4ng/&micro;L|| 0.2&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0&micro;L|| 0.1&micro;L|| 8.7&micro;L|| 10&micro;L
 |-
-|pT181attenuater 14:00 418.4ng/&micro;L|| 4.8&micro;L|| 0&micro;L|| 0&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 3&micro;L|| 0.3&micro;L|| 19.9&micro;L|| 30&micro;L
+|9/3 pT181attenuater 14:00 418.4ng/&micro;L|| 4.8&micro;L|| 0&micro;L|| 0&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 3&micro;L|| 0.3&micro;L|| 19.9&micro;L|| 30&micro;L
 |-
-|pT181attenuater 14:00 418.4ng/&micro;L|| 0.2&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.7&micro;L|| 10&micro;L
+|9/3 pT181attenuater 14:00 418.4ng/&micro;L|| 0.2&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.7&micro;L|| 10&micro;L
 |}
 </div>
@@ Line 82: / Line 82: @@
 ! ||DNA||EcoR1||Spe1||Xba1||Pst1||10xBuffer B(EcoR1&Spe1)||10xBuffer D(Xba1&Pst1)||100xBSA||MilliQ||total
 |-
-|pT181antisense 14:00 362.2ng/&micro;L|| 5.5&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 0&micro;L|| 3&micro;L|| 0&micro;L|| 0.3&micro;L|| 19.2&micro;L|| 30&micro;L
+|9/3 pT181antisense 14:00 362.2ng/&micro;L|| 5.5&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 0&micro;L|| 3&micro;L|| 0&micro;L|| 0.3&micro;L|| 19.2&micro;L|| 30&micro;L
 |-
-|pT181attenuater 14:00 418.4ng/&micro;L|| 0.3&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0&micro;L|| 0.1&micro;L|| 8.6&micro;L|| 10&micro;L
+|9/3 pT181attenuater 14:00 418.4ng/&micro;L|| 0.3&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0&micro;L|| 0.1&micro;L|| 8.6&micro;L|| 10&micro;L
 |-
-|pT181attenuater 14:00 418.4ng/&micro;L|| 5.5&micro;L|| 0&micro;L|| 0&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 3&micro;L|| 0.3&micro;L|| 19.2&micro;L|| 30&micro;L
+|9/3 pT181attenuater 14:00 418.4ng/&micro;L|| 5.5&micro;L|| 0&micro;L|| 0&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 3&micro;L|| 0.3&micro;L|| 19.2&micro;L|| 30&micro;L
 |-
-|pT181attenuater 14:00 418.4ng/&micro;L|| 0.3&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.6&micro;L|| 10&micro;L
+|9/3 pT181attenuater 14:00 418.4ng/&micro;L|| 0.3&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.6&micro;L|| 10&micro;L
 |}
 </div>
@@ Line 96: / Line 96: @@
 ! ||DNA||EcoR1||Spe1||10xBuffer B||100xBSA||MilliQ||total
 |-
-|apt 12_1R 14:00 507.5ng/&micro;L|| 3.9&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 3&micro;L|| 0.3&micro;L|| 20.8&micro;L|| 30&micro;L
+|9/3 apt 12_1R 14:00 507.5ng/&micro;L|| 3.9&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 3&micro;L|| 0.3&micro;L|| 20.8&micro;L|| 30&micro;L
 |-
-|apt 12_1R 14:00 507.5ng/&micro;L|| 0.2&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.7&micro;L|| 10&micro;L
+|9/3 apt 12_1R 14:00 507.5ng/&micro;L|| 0.2&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.7&micro;L|| 10&micro;L
 |}
 </div>
@@ Line 105: / Line 105: @@
 ! ||DNA||EcoR1||Xba1||10xBuffer D||100xBSA||MilliQ||total
 |-
-|DT 166.2ng/&micro;L|| 12&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 3&micro;L|| 0.3&micro;L|| 12.7&micro;L|| 30&micro;L
+|8/29 DT 166.2ng/&micro;L|| 12&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 3&micro;L|| 0.3&micro;L|| 12.7&micro;L|| 30&micro;L
 |-
-|DT 166.2ng/&micro;L|| 0.6&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.3&micro;L|| 10&micro;L
+|8/29 DT 166.2ng/&micro;L|| 0.6&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.3&micro;L|| 10&micro;L
+|}
+</div>
+<div class="experiment">
+{| class="wikitable"
+! ||DNA||EcoR1||Spe1||10xBuffer B||100xBSA||MilliQ||total
+|-
+|8/22 RBS-lysis1(2) 96ng/&micro;L|| 21&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 3&micro;L|| 0.3&micro;L|| 3.7&micro;L|| 30&micro;L
+|-
+|8/22 RBS-lysis1(2) 96ng/&micro;L|| 1.0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 7.9&micro;L|| 10&micro;L
+|}
+</div>
+<div class="experiment">
+{| class="wikitable"
+! ||DNA||EcoR1||Spe1||10xBuffer B||100xBSA||MilliQ||total
+|-
+|8/30 RBS-lysis2 220ng/&micro;L|| 9.1&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 3&micro;L|| 0.3&micro;L|| 15.6&micro;L|| 30&micro;L
+|-
+|8/30 RBS-lysis2 220ng/&micro;L|| 0.5&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.4&micro;L|| 10&micro;L
+|}
+</div>
+<div class="experiment">
+{| class="wikitable"
+! ||DNA||EcoR1||Spe1||10xBuffer B||100xBSA||MilliQ||total
+|-
+|8/20 RBS-lysis3(1) 282ng/&micro;L|| 7.1&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 3&micro;L|| 0.3&micro;L|| 17.6&micro;L|| 30&micro;L
+|-
+|8/20 RBS-lysis3(1) 282ng/&micro;L|| 0.4&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.5&micro;L|| 10&micro;L
 |}
 </div>

Template:Kyoto/Notebook/Sep 3

From 2013.igem.org

Revision as of 10:37, 25 September 2013

Contents

Sep 3

Electrophoresis

Miniprep

Liquid Culture

Restriction Enzyme Digestion

PCR

Mutatation PCR

Colony PCR

Restriction Enzyme Digestion

Gel Extraction

Liquid Culture