From 2014.igem.org

(Difference between revisions)

Revision as of 07:25, 23 September 2014

count down

アメフラシノート

日付

Liquid Culture

名前

Sample	Medium
サンプル名	培地

日付

Gel Extraction

名前

Lane	Sample
レーン番号	サンプル名

日付

Electrophoresis

名前

Lane	Sample	Ensyme1	Ensyme2
レーン番号	サンプル名	制限酵素1	制限酵素2

日付

Miniprep/DNA Purification

名前

Sample	Concentration(µg/mL)	260/280	260/230
サンプル名	(Concentration)	(260/280)	(260/280)

日付

Restriction Enzyme Digestion

名前

Sample		EcoR1	Xba1	Spe1	Pst1	Buffer		BSA	MilliQ	Total
サンプル名	(サンプル名)	(E)	(X)	(S)	(P)	(HorM)	(Buffer)	(BSA)	(MilliQ)	(Total)

日付

Ligation

名前

Vector		Insert		Buffer		Ligase		MilliQ	Total
ベクター名	(ベクター)	インサート名	(インサート)	(HorM)	(Buffer)	Ligase名	(Ligase)	(MilliQ)	(Total)

日付

Transformation

名前

Sample		CompetentCells		Total	Medium
サンプル名	(サンプル)	コンピ名	(コンピ)	(Total)	培地

日付

Dephosphorylation

名前

Sample
サンプル名	(サンプル)

7/31

Restriction Enzyme Digestion

Nakamura Matsumoto

Sample	Volume	EcoR1	Spe1	Buffer(H)	MilliQ	TOTAL
3/24 PCR(dddD)cloning	3.2	1	1	3	21.8	30
pSB1C3	18.5	1	1	3	6.5	30
pSB1C3(control)	6.2	0	0	0	2.8	10

8/1

Gel Extraction

Yasuda Murata

Lane	DNA
1	1kbp ladder
2	3/24 PCR(dddD)cloning E,S
3	3/24 PCR(dddD)cloning E,S
4	Blank
5	pSB1C3
6	pSB1C3
7	Blank
8	1kbp ladder
9	3/24 PCR(dddD)cloning(control)
10	pSB1C3(control)

8/1

Ligation

Yasuda

Vector	inserter	10xt4LigaseBuffer	t4Ligase	MilliQ	TOTAL
1.5 µL(8/1 pSB1C3 2047bp 13.3µg/mL 20μg)	6.9 µL(8/1 dddD 2545bp 10.8µg/mL 74.60µg)	2µL	1µL	8.6µL	20µL

8/1

transformation

Murata

Name	Sample/(µl)	Competent Cells/(µl)	Total/(µl)	Plate
pSB1C3	3	30	165	SOC(132µl)

8/2

PCR Cloning

Murata

	dddD 121µg/µL
2x KAPA HiFiHotStartReadyMix	12.5µL
F Pre-dddD cloning 62	0.75µL
Rv Spe1-dddDcloning 68	0.75µL
MilliQ	11µL
TOTAL	25µL

8/2

Colony PCR

Honda Matsumoto Nakamura

	Master Mix
Quick Taq	15µL
VF2	0.6µL
VR	0.6µL
MilliQ	13.8µL
TOTAL	30µL

8/2

Restriction Enzyme Digestion

Nakamura Matsumoto

Sample	Volume	EcoR1	Spe1	Buffer	MilliQ	TOTAL
8/2 dddD cloning	8.2	1	1	3	16.8	30
8/2 dddD cloning control	0.41	0	0	1	8.6	10
pSB1C3(control)	6.2	0	0	3(H)	2.8	10

8/3

Electrophoresis for confirmation

Murata

Lane	Sample
1	pSB1C3 dddD 1
2	pSB1C3 dddD 2
3	pSB1C3 dddD 3
4	1kb ladder(3µL)
5	1kb ladder(1µL)

8/3

Gel Extraction

Tatsui

Lane	DNA
1	1kbp ladder
2	Blank
3	pSB1C3 25µμL 2070bp
4	Blank
5	pSB1C3 5µL
6	Blank
7	8/2 cloning dddD 25µL 2544bp
8	Blank
9	3/24 PCR(dddD)cloning(control)
10	8/2 cloning dddD 5µL

9/14

Gel Extraction

Honda,Yasuda

Lane	DNA
1	1kbp ladder
2	3/24 PCR(dddD)cloning E,S
3	9/13SAMmT(control)
4	9/13SAMmT(EP)
5	9/13Pcon-pSB1C3(control)
6	9/13Pcon-pSB1C3(XS)

9/13

Column cleanup

Yasuda

Sample	density(µg/mL)	260/280	260/230
9/12 AT-EP	2.3	1.33	1.11
9/12 AT-His-XP	2.0	1.33	0.86

9/13

Ligation

Honda

sumple	Vector	Insert	Ligase	2xBuffer	MilliQ
9/13 column cleanup AT(EP)+9/8 pSB1C3(EP)	1	3	1	5	0
9/13 column cleanup AT-His(XP)+9/13 pSB1C3(XP)	1	3	1	5	0

9/13

Miniprep

Honda

Sample	density(µg/mL)	260/280	260/230
9/12 SAMmt-YCPlac111	140	1.80	1.90
9/12 T7-RBS-psB1C3	290	1.78	1.65

9/13

Liquid Culture

Yasuda

Sample	Medium
9/12 dddD-pSB1C3	CP
9/12 dddD-pSB1C3	CP
9/12 dddD-pSB1C3	CP
9/12 T7-RBS-His	CP
9/12 T7-RBS-His	CP
9/12 T7-RBS-His	CP
control	CP

9/14

Colony PCR

Yasuda

	Master Mix
Quick Taq	335
VF	13.4
VR	13.4
MilliQ	308.2
TOTAL	670

9/14

Gel Extraction

Honda

Lane	DNA	volume
1	1kbp ladder	5
2	3/24 PCR(dddD)cloning E,S	15
3	9/13SAMmT(control)	15
4	9/13SAMmT(EP)	15
5	9/13Pcon-pSB1C3(control)	15
6	9/13Pcon-pSB1C3(XS)

9/14

Electrophoresis for confirmation

Honda, Li and Yamauchi

Lane	Restriction Enzyme Digestion Products
1	9/12 AT-YcPlac 1~8
2	9/12 AT-YcPlac 1~8
3	9/12 AT-YcPlac 1~8
4	9/12 AT-YcPlac 1~8
5	9/12 AT-YcPlac 1~8
6	9/12 AT-YcPlac 1~8
7	9/12 AT-YcPlac 1~8
8	9/12 AT-YcPlac 1~8
9	GeneRuler
10	9/12 AT-His-YcPlac 1~8
11	9/12 AT-His-YcPlac 1~8
12	9/12 AT-His-YcPlac 1~8
13	9/12 AT-His-YcPlac 1~8
14	9/12 AT-His-YcPlac 1~8
15	9/12 AT-His-YcPlac 1~8
16	9/12 AT-His-YcPlac 1~8
17	9/12 AT-His-YcPlac 1~8

Lane	Restriction Enzyme Digestion Products
1	9/12 mamB-YcPlac 1~8
2	9/12 mamB-YcPlac 1~8
3	9/12 mamB-YcPlac 1~8
4	9/12 mamB-YcPlac 1~8
5	9/12 mamB-YcPlac 1~8
6	9/12 mamB-YcPlac 1~8
7	9/12 mamB-YcPlac 1~8
8	9/12 mamB-YcPlac 1~8
9	GeneRuler
10	9/12 DECARB-YcPlac 1~8
11	9/12 DECARB-YcPlac 1~8
12	9/12 DECARB-YcPlac 1~8
13	9/12 DECARB-YcPlac 1~8
14	9/12 DECARB-YcPlac 1~8
15	9/12 DECARB-YcPlac 1~8
16	9/12 DECARB-YcPlac 1~8
17	9/12 DECARB-YcPlac 1~8

Lane	Restriction Enzyme Digestion Products
1	9/12 DECARB 9~16
2	9/12 DECARB 9~16
3	9/12 DECARB 9~16
4	9/12 DECARB 9~16
5	9/12 DECARB 9~16
6	9/12 DECARB 9~16
7	9/12 DECARB 9~16
8	9/12 DECARB 9~16
9	GeneRuler-YcPlac
10	9/12 DECARB-YcPlac 17~20
11	9/12 DECARB-Ycplac 17~20
12	9/12 DECARB-Ycplac 17~20
13	9/12 DiDECARB-YcPlac 1~4
14	9/12 DiDECARB-YcPlac 1~4
15	9/12 DiDECARB-YcPlac 1~4
16	9/12 DiDECARB-YcPlac 1~4
17	9/12 DiDECARB-YcPlac 1~4

Lane	Restriction Enzyme Digestion Products
1	9/12 DiDECARB-YcPlac 5~12
2	9/12 DiDECARB-YcPlac 5~12
3	9/12 DiDECARB-YcPlac 5~12
4	9/12 DiDECARB-YcPlac 5~12
5	9/12 DiDECARB-YcPlac 5~12
6	9/12 DiDECARB-YcPlac 5~12
7	9/12 DiDECARB-YcPlac 5~12
8	9/12 DiDECARB-YcPlac 5~12
9	GeneRuler
10	9/12 DiDECARB-YcPlac 13~20
11	9/12 DiDECARB-YcPlac 13~20
12	9/12 DiDECARB-YcPlac 13~20
13	9/12 DiDECARB-YcPlac 13~20
14	9/12 DiDECARB-YcPlac 13~20
15	9/12 DiDECARB-YcPlac 13~20
16	9/12 DiDECARB-YcPlac 13~20
17	9/12 DiDECARB-YcPlac 13~20

9/14

Miniprep

Honda

DNA	density(µg/ml)	260/280	260/230
9/14 dddD(µ)-pSB1C3	645	1.82	1.70
9/14 dddD(µ)-pSB1C3	660	1.69	1.74
9/14 dddD(µ)-pSB1C3	635	1.87	1.57
9/14 T7-RBS-His-pSB1C3	440	1.65	1.23
9/14 T7-RBS-His-pSB1C3	450	1.92	1.46
9/14 T7-RBS-His-pSB1C3	190	1.ss	1.26

9/14

Restriction Enzyme Digestion

Honda

Sample	Volume	Spe1	Pst1	Xba1)	Buffur	BSA	MilliQ	TOTAL
9/14 dddD(µ)-pSB1C3	6	0	2	2	6(M)	6	38	60
9/14 dddD(µ)-pSB1C3	6	0	2	2	6(M)	6	38	60
9/14 dddD(µ)-pSB1C3	6	0	2	2	6(M)	6	38	60
9/14 T7-RBS-His-pSB1C3	5	1	1	0	3(H)	0	18	30
9/14 T7-RBS-His-pSB1C3	5	1	1	0	3(H)	0	18	30
9/14 T7-RBS-His-pSB1C3	10	1	1	0	3(H)	0	15	30

9/14

Ligation

Yasuda

NEB T4DNALigase	10XLig Buffer	9/12 DECARB XS column	9/14 Pcon-pSB1C3 XS Dephosphorylated	MilliQ	TOTAL
1	2	2.93	0.83	13.24	20

NEB T4DNALigase	10XLig Buffer	9/12 DiDECARB XS column	9/14 Pcon-pSB1C3 XS Dephosphorylated	MilliQ	TOTAL
1	2	3.66	0.83	12.5	20

@@ Line 13: / Line 13: @@
-<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”>
+<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”></a>
 <h3>日付</h3>
 <h4>Liquid Culture</h4>
@@ Line 23: / Line 23: @@
-<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”>
+<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”></a>
 <h3>日付</h3>
 <h4>Gel Extraction</h4>
@@ Line 33: / Line 33: @@
-<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”>
+<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”></a>
 <h3>日付</h3>
 <h4>Electrophoresis</h4>
@@ Line 43: / Line 43: @@
-<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”>
+<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”></a>
 <h3>日付</h3>
 <h4>Miniprep/DNA Purification</h4>
@@ Line 53: / Line 53: @@
-<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”>
+<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”></a>
 <h3>日付</h3>
 <h4>Restriction Enzyme Digestion</h4>
@@ Line 63: / Line 63: @@
-<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”>
+<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”></a>
 <h3>日付</h3>
 <h4>Ligation</h4>
@@ Line 73: / Line 73: @@
-<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”>
+<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”></a>
 <h3>日付</h3>
 <h4>Transformation</h4>
@@ Line 83: / Line 83: @@
-<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”>
+<a name=“半角小文字日付（ex. September23）” class=“kyoto-jump”></a>
 <h3>日付</h3>
 <h4>Dephosphorylation</h4>

Team:Kyoto/Notebook/DMS Synthesis

From 2014.igem.org

Revision as of 07:25, 23 September 2014

アメフラシノート

日付

Liquid Culture

日付

Gel Extraction

日付

Electrophoresis

日付

Miniprep/DNA Purification

日付

Restriction Enzyme Digestion

日付

Ligation

日付

Transformation

日付

Dephosphorylation

7/31

Restriction Enzyme Digestion

8/1

Gel Extraction

8/1

Ligation

8/1

transformation

8/2

PCR Cloning

8/2

Colony PCR

8/2

Restriction Enzyme Digestion

8/3

Electrophoresis for confirmation

8/3

Gel Extraction

9/14

Gel Extraction

9/13

Column cleanup

9/13

Ligation

9/13

Miniprep

9/13

Liquid Culture

9/14

Colony PCR

9/14

Gel Extraction

9/14

Electrophoresis for confirmation

9/14

Miniprep

9/14

Restriction Enzyme Digestion

9/14

Ligation