Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)

Revision as of 07:33, 24 September 2013

Sep 6

Liquid Culture

No name

Sample	medium
9/4 RBS-lysis3＋DT3	Plusgrow medium(+CP)
9/4 RBS-lysis1＋DT3	Plusgrow medium(+CP)
9/4 psB4K5	Plusgrow medium(+K)

Colony PCR

Nakamoto

Sample	base pair
8/15 BBa_K112806(lysis2) -(1)	828
negative control	--

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

Sample	base pair
8/10 BBa_Q0400 -(1)	3995
negative control	--

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

Miniprep

No name

DNA	concentration[µg/mL]	260/280	260/230
(・▽・)

Restriction Enzyme Digestion

Murata, Murakami, Nakamoto, Ashida, Matsumoto and Yoshida

	8/11 J23100 -(2)	SpeI	PstI	Buffer	MilliQ	total
2 cuts	4.0µL	1.0µL	1.0µL	3.0µL	21µL	30µL
1 cut	1.3µL	0.2µL	--	1.0µL	7.5µL	10µL
1 cut	1.3µL	--	0.2µL	1.0µL	7.5µL	10µL
NC	1.3µL	--	--	1.0µL	7.7µL	10µL

at 37°C, for 1h

	8/15 B0034(RBS) -(2)	SpeI	PstI	Buffer	MilliQ	total
2 cuts	6.0µL	1.0µL	1.0µL	3.0µL	19µL	30µL
1 cut	2.0µL	0.2µL	--	1.0µL	6.8µL	10µL
1 cut	2.0µL	--	0.2µL	1.0µL	6.8µL	10µL
NC	2.0µL	--	--	1.0µL	7.0µL	10µL

at 37°C, for 1h

	8/17 K117000(lysis1) -(1)	XbaI	PstI	BSA	Buffer	MilliQ	total
2 cuts	2.0µL	1.0µL	1.0µL	3.0µL	3.0µL	20µL	30µL
1 cut	1.0µL	0.2µL	--	1.0µL	1.0µL	6.8µL	10µL
1 cut	1.0µL	--	0.2µL	1.0µL	1.0µL	6.8µL	10µL
NC	1.0µL	--	--	1.0µL	1.0µL	7.0µL	10µL

at 37°C, for 1h

	8/17 K748002(lysis3) -(1)	XbaI	PstI	BSA	Buffer	MilliQ	total
2 cuts	2.0µL	1.0µL	1.0µL	3.0µL	3.0µL	20µL	30µL
1 cut	1.0µL	0.2µL	--	1.0µL	1.0µL	6.8µL	10µL
1 cut	1.0µL	--	0.2µL	1.0µL	1.0µL	6.8µL	10µL
NC	1.0µL	--	--	1.0µL	1.0µL	7.0µL	10µL

at 37°C, for 1h

	8/17 I13504(RBS-GFP-DT) -(1)	XbaI	PstI	BSA	Buffer	MilliQ	total
2 cuts	2.0µL	1.0µL	1.0µL	3.0µL	3.0µL	20µL	30µL
1 cut	1.0µL	0.2µL	--	1.0µL	1.0µL	6.8µL	10µL
1 cut	1.0µL	--	0.2µL	1.0µL	1.0µL	6.8µL	10µL
NC	1.0µL	--	--	1.0µL	1.0µL	7.0µL	10µL

at 37°C, for 1h

	8/17 I0462(RBS-luxR-DT) -(1)	XbaI	PstI	BSA	Buffer	MilliQ	total
2 cuts	2.0µL	1.0µL	1.0µL	3.0µL	3.0µL	20µL	30µL
1 cut	1.0µL	0.2µL	--	1.0µL	1.0µL	6.8µL	10µL
1 cut	1.0µL	--	0.2µL	1.0µL	1.0µL	6.8µL	10µL
NC	1.0µL	--	--	1.0µL	1.0µL	7.0µL	10µL

at 37°C, for 1h

	8/17 I732020(RBS-lacZα-DT) -(1)	XbaI	PstI	BSA	Buffer	MilliQ	total
2 cuts	2.0µL	1.0µL	1.0µL	3.0µL	3.0µL	20µL	30µL
1 cut	1.0µL	0.2µL	--	1.0µL	1.0µL	6.8µL	10µL
1 cut	1.0µL	--	0.2µL	1.0µL	1.0µL	6.8µL	10µL
NC	1.0µL	--	--	1.0µL	1.0µL	7.0µL	10µL

at 37°C, for 1h

	8/11 R6061 -(1)	SpeI	PstI	Buffer	MilliQ	total
2 cuts	8.0µL	1.0µL	1.0µL	3.0µL	17µL	30µL
1 cut	3.0µL	0.2µL	--	1.0µL	5.8µL	10µL
1 cut	3.0µL	--	0.2µL	1.0µL	5.8µL	10µL
NC	3.0µL	--	--	1.0µL	6.0µL	10µL

at 37°C, for 1h

Gel Extraction

Ashida

Lane	DNA	Enzyme
1	100bp ladder	--
3	8/18 K117000(lysis2) -1	XbaI & PstI
5	8/18 K117000(lysis2) -1	XbaI & PstI

File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4

Name	concentration[µg/mL]	260/280	260/230
8/18 lysis2-1(XbaI & PstI)	3.1	3.64	0.19
8/18 lysis2-1(XbaI & PstI)	3.4	1.89	0.13

Liquid Culture

Nakamoto

Sample	medium
8/15 BBa_K112806(lysis2) -(1)	Plusgrow medium(+CP)
8/15 BBa_Q04400(tetR) -(1)	Plusgrow medium(+Amp)

Colony PCR

Nakamoto

Sample	base pair
8/15 BBa_K112806(lysis2) -(1)	828
negative control	--

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

Sample	base pair
8/10 BBa_Q0400 -(1)	3995
negative control	--

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

Colony PCR

Nakamoto

Sample	base pair
8/15 BBa_K112806(lysis2) -(1)	828
negative control	--

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

Sample	base pair
8/10 BBa_Q0400 -(1)	3995
negative control	--

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

@@ Line 1: / Line 1: @@
 ==Sep 6==
-===Colony PCR===
-<div class="experiment">
-<span class="author">Nakamoto</span>
-{| class="wikitable"
-!|Sample||base pair
-|-
-|8/15 BBa_K112806(lysis2) -(1)||828
-|-
-|negative control||--
-|}
-{| class="wikitable"
-!PreDenature||Denature||Annealing||Extension||cycle
-|-
-|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
-|-
-|5min||30s||30s||1min||30cycles
-|}
-{| class="wikitable"
-!Sample||base pair
-|-
-|8/10 BBa_Q0400 -(1)||3995
-|-
-|negative control||--
-|}
-{| class="wikitable"
-!PreDenature||Denature||Annealing||Extension||cycle
-|-
-|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
-|-
-|5min||30s||30s||30s||30cycles
-|}
 ===Liquid Culture===
 <!-- ここから -->
@@ Line 39: / Line 7: @@
 !Sample||medium
 |-
-|(・ω・) ||
+|9/4 RBS-lysis3＋DT3 ||Plusgrow medium(+CP)
+|-
+|9/4 RBS-lysis1＋DT3 ||　Plusgrow medium(+CP)
 |-
-|（・Ｘ・） ||
+|9/4 psB4K5||Plusgrow medium(+K)
 |}
 </div>
 ===Colony PCR===

Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

Revision as of 07:33, 24 September 2013

Contents

Sep 6

Liquid Culture

Colony PCR

Miniprep

Restriction Enzyme Digestion

Gel Extraction

Liquid Culture

Colony PCR

Colony PCR