Template:Kyoto/Notebook/Sep 3

From 2013.igem.org

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Revision as of 12:27, 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

PCR

Hirano

Plac(1) plasmid 153.7ng/µL	10x buffer KOD plus-ver.2	2mM dNTPs	25mM MgSO4	primer (KaiA_BsaI_Mut_fwd)	primer (KaiA_BsaI_Mut_rer)	KOD plus-ver.2	MilliQ	total
0.13	2.5	2.5	1.5	0.75	0.75	0.5	16.37	25

KaiB (plasmid) 20.2ng/µL	10x buffer KOD plus-ver.2	2mM dNTPs	25mM MgSO4	primer (GG2_RBS_KaiB_fwd)	primer (KaiB_GG3_rer)	KOD plus-ver.2	MilliQ	total
1	2.5	2.5	1.5	0.75	0.75	0.5	15.5	25

KaiC (plasmid) 4.4ng/µL	10x buffer KOD plus-ver.2	2mM dNTPs	25mM MgSO4	primer	primer	KOD plus-ver.2	MilliQ	total
1	2.5	2.5	1.5	0.75	0.75	0.5	15.5	25

DT-1 (plasmid) 153.7ng/µL	10x buffer KOD plus-ver.2	2mM dNTPs	25mM MgSO4	primer	primer	KOD plus-ver.2	MilliQ	total
0.1	2.5	2.5	1.5	0.75	0.75	0.5	16.4	25

PKaiBC(PCR product) 16.3ng/µL	10x buffer KOD plus-ver.2	2mM dNTPs	25mM MgSO4	primer	primer	KOD plus-ver.2	MilliQ	total
0.7	2.5	2.5	1.5	0.75	0.75	0.5	15.8	25

SasA(PCR product) 21.6ng/µL	10x buffer KOD plus-ver.2	2mM dNTPs	25mM MgSO4	primer	primer	KOD plus-ver.2	MilliQ	total
0.5	2.5	2.5	1.5	0.75	0.75	0.5	16	25

RpaA(PCR product) 15.4ng/µL	10x buffer KOD plus-ver.2	2mM dNTPs	25mM MgSO4	primer	primer	KOD plus-ver.2	MilliQ	total
0.6	2.5	2.5	1.5	0.75	0.75	0.5	15.9	25

RpaB(PCR product) 26.7ng/µL	10x buffer KOD plus-ver.2	2mM dNTPs	25mM MgSO4	primer	primer	KOD plus-ver.2	MilliQ	total
0.4	2.5	2.5	1.5	0.75	0.75	0.5	16.1	25

@@ Line 146: / Line 146: @@
 <span class="author">Hirano</span>
 {| class="wikitable"
-!Plac(1) plasmid 153.7ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer (KaiA_BsaI_Mut_fwd)||primer (KaiA_BsaI_Mut_rer||KOD plus-ver.2||MilliQ||total
+!Plac(1) plasmid 153.7ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer (KaiA_BsaI_Mut_fwd)||primer (KaiA_BsaI_Mut_rer)||KOD plus-ver.2||MilliQ||total
 |-
 |0.13 ||2.5||2.5||1.5||0.75||0.75||0.5||16.37||25
@@ Line 153: / Line 153: @@
 <div class="experiment">
 {| class="wikitable"
-!KaiB (plasmid) 20.2ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer (GG2_RBS_KaiB_fwd)||primer (KaiB_GG3_rer||KOD plus-ver.2||MilliQ||total
+!KaiB (plasmid) 20.2ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer (GG2_RBS_KaiB_fwd)||primer (KaiB_GG3_rer)||KOD plus-ver.2||MilliQ||total
 |-
 |1||2.5||2.5||1.5||0.75||0.75||0.5||15.5||25
+|}
+</div>
+<div class="experiment">
+{| class="wikitable"
+!KaiC (plasmid) 4.4ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer ||primer ||KOD plus-ver.2||MilliQ||total
+|-
+|1||2.5||2.5||1.5||0.75||0.75||0.5||15.5||25
+|}
+</div>
+<div class="experiment">
+{| class="wikitable"
+!DT-1 (plasmid) 153.7ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer ||primer ||KOD plus-ver.2||MilliQ||total
+|-
+|0.1||2.5||2.5||1.5||0.75||0.75||0.5||16.4||25
+|}
+</div>
+<div class="experiment">
+{| class="wikitable"
+!PKaiBC(PCR product) 16.3ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer ||primer ||KOD plus-ver.2||MilliQ||total
+|-
+|0.7||2.5||2.5||1.5||0.75||0.75||0.5||15.8||25
+|}
+</div>
+<div class="experiment">
+{| class="wikitable"
+!SasA(PCR product) 21.6ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer ||primer ||KOD plus-ver.2||MilliQ||total
+|-
+|0.5||2.5||2.5||1.5||0.75||0.75||0.5||16||25
+|}
+</div>
+<div class="experiment">
+{| class="wikitable"
+!RpaA(PCR product) 15.4ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer ||primer ||KOD plus-ver.2||MilliQ||total
+|-
+|0.6||2.5||2.5||1.5||0.75||0.75||0.5||15.9||25
+|}
+</div>
+<div class="experiment">
+{| class="wikitable"
+!RpaB(PCR product) 26.7ng/&micro;L||10x buffer KOD plus-ver.2|||2mM dNTPs||25mM MgSO4||primer ||primer ||KOD plus-ver.2||MilliQ||total
+|-
+|0.4||2.5||2.5||1.5||0.75||0.75||0.5||16.1||25
 |}
 </div>

Template:Kyoto/Notebook/Sep 3

From 2013.igem.org

Revision as of 12:27, 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