Template:Kyoto/Notebook/Sep 7

From 2013.igem.org

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Revision as of 13:36, 24 September 2013

Sep 7

Gel Extraction

No Name

Name	concentration[µg/mL]	260/280	260/230
apt 12_1R-DT	12.7	1.62	5.74
RBS-lysis2-DT	6.7	1.71	5.46
Pλ-luvI	8.1	1.76	2.25

Miniprep

No name

DNA	concentration[µg/mL]	260/280	260/230
RBS-lysis1-DT	195.3
RBS-lysis3-DT	255.5	1.91	2.04
Ptet-pT18	183.6	1.85	1.48
Pcon-RBS-tetR-DT	134.3	1.94	2.20
Ptet-RBS-lacZα-DT	67.7	1.92	1.79
Pcon-pT181 attenuator	232.3	1.95	1.90
pSB4K5	237.0	1.61	1.50
pT181 antisense(pSB1C3)	256.4	1.63	1.78
apt12_1R(pSB1C3)	230.0	1.98	2.28

Ligation

No Name

state	Vector		Inserter		Ligation High ver.2
experiment	9/6 Plux (SpeI&PstI)	2 µL	9/7 RBS-lysis2-DT (EcoRI&XbaI)	µL	3.5 µL
experiment	9/6 Plac (SpeI&PstI)	µL	9/7 RBS-lysis2-DT (EcoRI&XbaI)	µL	3.5 µL
experiment	9/6 Pcon-RBS-luxR-DT (EcoRI&XbaI)	1.9 µL	9/6 Plux-RBS-GFP-DT(SpeI&PstI)	2.2 µL	3.5 µL
experiment	9/6 Pbed/araC (SpeI&PstI)	1.6 µL	9/5 RBS-luxI-DT (EcoRI&XbaI)	16 µL	3.5 µL
experiment	9/6 Plac (SpeI&PstI)	1.5 µL	9/5 pT181 antisense (EcoRI&XbaI)	2.0 µL	3.5 µL
experiment	9/6 Pcon (SpeI&PstI)	3.0 µL	9/5 RBS-luxI-DT (EcoRI&XbaI)	10 µL	3.5 µL

Colony PCR

No name

Sample	base pair
9/5 Plac-spinarch-DT-13~20
9/5 Plac-pT181 attenuator-13~20

PreDenature	Denature	Annealing	Extension	cycle
94　°C	94　°C	55　°C	60　°C	--
5min	30s	30s	42s	30cycle

Transformation

Tatsui

Name	Sample	Competent Cells	Total
Pcon-RBS-luxR-DT+Plux-RBS-GFP-DT	2µL	20µL	22µL
Pbed/araC+RBS-luxI-DT	2µL	20µL	22µL
Plac+pT181 antisence	2µL	20µL	22µL
Pcon+RBS-luxI-DT	2µL	20µL	22µL

Restriction Enzyme Digestion

No Name

	DNA	XbaI	PstI	Buffer	BSA	MilliQ	total
9/7 RBS-lysis1-DT(XbaI & PstI)	10 µL	1 µL	1 µL	3 µL	3 µL	12 µL	30 µL
NC	0.5 µL	0 µL	0 µL	1 µL	1 µL	7.5 µL	10 µL

	DNA	XbaI	PstI	Buffer	BSA	MilliQ	total
9/7 RBS-lysis3-DT (XbaI & PstI)	7.8 µL	1 µL	1 µL	3 µL	3 µL	14.2 µL	30 µL
NC	0.4 µL	0 µL	0 µL	1 µL	1 µL	7.6 µL	10 µL

	DNA	SpeI	PstI	Buffer	BSA	MilliQ	total
9/5 Plac (SpeI & PstI)	6.0 µL	1 µL	1 µL	3 µL	3 µL	16.0 µL	30 µL
NC	0.3 µL	0 µL	0 µL	1 µL	1 µL	7.7 µL	10 µL

	DNA	EcorI	SpeI	Buffer	BSA	MilliQ	total
9/7 pSB4K5 (EcoRI & SpeI)	8.4 µL	1 µL	1 µL	3 µL	3 µL	16.0 µL	30 µL
NC	0.4 µL	0 µL	0 µL	1 µL	1 µL	7.7 µL	10 µL

	DNA	restricion enzyme	restriction enzyme	Buffer	BSA	MilliQ	total
Pcon-pT181 attenuator (SpeI & PstI)	8.6 µL	1 µL	1 µL	3 µL	3 µL	13.4 µL	30 µL
Pcon-pT181 attenuator (EcorI & SpeI)	8.6 µL	1 µL	1 µL	3 µL	3 µL	13.4 µL	30 µL
NC	0.4 µL	0 µL	0 µL	1 µL	1 µL	7.6 µL	10 µL

	DNA	XbaI	PstI	Buffer	BSA	MilliQ	total
9/5 Spin-DT (XbaI & PstI)	5.4 µL	1 µL	1 µL	3 µL	3 µL	16.6 µL	30 µL
NC	0.3 µL	0 µL	0 µL	1 µL	1 µL	7.7 µL	10 µL

	DNA	SpeI	PstI	Buffer	BSA	MilliQ	total
Ptet-anticodone (SpeI & PstI)	11 µL	1 µL	1 µL	3 µL	3 µL	11 µL	30 µL
NC	0.5 µL	0 µL	0 µL	1 µL	1 µL	7.5 µL	10 µL

	DNA	XbaI	PstI	Buffer	BSA	MilliQ	total
9/6 RBS-lacZα-DT (XbaI & PstI)	11 µL	1 µL	1 µL	3 µL	3 µL	11 µL	30 µL
NC	0.5 µL	0 µL	0 µL	1 µL	1 µL	7.5 µL	10 µL

	DNA	EcoRI	SpeI	Buffer	BSA	MilliQ	total
pSB1C3 (1) (EcoRI & SpeI)	6.6 µL	1 µL	1 µL	3 µL	3 µL	15.4 µL	30 µL
NC	0.3 µL	0 µL	0 µL	1 µL	1 µL	7.7 µL	10 µL

	DNA	XbaI	PstI	Buffer	BSA	MilliQ	total
pSB1C3 (3) (XbaI & PstI)	6.6 µL	1 µL	1 µL	3 µL	3 µL	15.4 µL	30 µL
NC	0.3 µL	0 µL	0 µL	1 µL	1 µL	7.7 µL	10 µL

@@ Line 110: / Line 110: @@
 |-
 |NC||0.5 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.5 &micro;L||10 &micro;L
+|}
+<br>
+{| class="wikitable"
 ! ||DNA||XbaI||PstI||Buffer||BSA||MilliQ||total
 |-
@@ Line 115: / Line 118: @@
 |-
 |NC||0.4 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.6 &micro;L||10 &micro;L
+|}
 <br>
+{| class="wikitable"
 ! ||DNA||SpeI||PstI||Buffer||BSA||MilliQ||total
 |-
@@ Line 121: / Line 126: @@
 |-
 |NC||0.3 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.7 &micro;L||10 &micro;L
+|}
 <br>
+{| class="wikitable"
 ! ||DNA||EcorI||SpeI||Buffer||BSA||MilliQ||total
 |-
@@ Line 127: / Line 134: @@
 |-
 |NC||0.4 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.7 &micro;L||10 &micro;L
+|}
 <br>
+{| class="wikitable"
 ! ||DNA||restricion enzyme||restriction enzyme||Buffer||BSA||MilliQ||total
 |-
@@ Line 135: / Line 144: @@
 |-
 |NC||0.4 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.6 &micro;L||10 &micro;L
+|}
 <br>
+{| class="wikitable"
 ! ||DNA||XbaI||PstI||Buffer||BSA||MilliQ||total
 |-
@@ Line 141: / Line 152: @@
 |-
 |NC||0.3 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.7 &micro;L||10 &micro;L
+|}
 <br>
+{| class="wikitable"
 ! ||DNA||SpeI||PstI||Buffer||BSA||MilliQ||total
 |-
@@ Line 147: / Line 160: @@
 |-
 |NC||0.5 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.5 &micro;L||10 &micro;L
+|}
 <br>
+{| class="wikitable"
 ! ||DNA||XbaI||PstI||Buffer||BSA||MilliQ||total
 |-
@@ Line 153: / Line 168: @@
 |-
 |NC||0.5 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.5 &micro;L||10 &micro;L
+|}
 <br>
+{| class="wikitable"
 ! ||DNA||EcoRI||SpeI||Buffer||BSA||MilliQ||total
 |-
@@ Line 159: / Line 176: @@
 |-
 |NC||0.3 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.7 &micro;L||10 &micro;L
+|}
 <br>
+{| class="wikitable"
 ! ||DNA||XbaI||PstI||Buffer||BSA||MilliQ||total
 |-

Template:Kyoto/Notebook/Sep 7

From 2013.igem.org

Revision as of 13:36, 24 September 2013

Contents

Sep 7

Gel Extraction

Miniprep

Ligation

Colony PCR

Transformation

Restriction Enzyme Digestion