Template:Kyoto/Notebook/Sep 9

From 2013.igem.org

(Difference between revisions)

Revision as of 04:49, 24 September 2013

Sep 9

PCR

Nakagawa

KaiA(plasmid)	10x buffer	dNTPs	MgSO4	Primer(661-RBS-KaiA-fwd)	Primer(kaiA-662-rev-ver2)	Kod-plus ver2	MilliQ	total
1.0	2.5	2.5	1.5	0.75	0.75	0.5	15.5	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	58.5°C	68°C	--
2min	10s	30s	30s	30cycles

KaiB(plasmid)	10x buffer	dNTPs	MgSO4	Primer(661-RBS-KaiA-fwd-ver2)	Primer(kaiB-663-rev-ver2)	Kod-plus ver2	MilliQ	total
1.0	2.5	2.5	1.5	0.75	0.75	0.5	15.5	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	58.5°C	68°C	--
2min	10s	30s	30s	30cycles

P-KaiBC(PCR products)	10x buffer	dNTPs	MgSO4	Primer(prefix-P-kaiBC-fwd-ver2)	Primer(P-KaiBC-suffix-rev-verw)	Kod-plus ver2	MilliQ	total
0.7	2.5	2.5	1.5	0.75	0.75	0.5	15.8	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	58.5°C	68°C	--
2min	10s	30s	30s	30cycles

DT(plasmid)	10x buffer	dNTPs	MgSO4	Primer(664-DT-fwd-ver2)	Primer(DT-suffix-660-rev-ver2)	Kod-plus ver2	MilliQ	total
1.0	2.5	2.5	1.5	0.75	0.75	0.5	15.5	25

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	58.5°C	68°C	--
2min	10s	30s	30s	30cycles

Miniprep

No name

DNA	concentration[µg/mL]	260/280	260/230
9/8 pSB1C3	83.2	1.45	0.76
9/8 RBS-lysis2-DT	72.8	1.67	0.97
9/8 spinach-DT	152.6	1.88	1.97
Nuclease-Free water	-0.4	-14.30	0.25

Restriction Enzyme Digestion

じっけんしゃ

	DNA	restricion enzyme	restriction enzyme	Buffer	BSA	MilliQ	total
2 cuts	（°°） µL	µL	µL	µL	µL	µL	µL
NC	µL	µL	µL	µL	µL	µL	µL

Transformation

じっけんしゃ

Name	Sample	Competent Cells	Total	Plate
いでんし	なんちゃらµL	ごにょごにょµL	ほにゃららµL	こーせーぶっしつ
いかどうよう

Ligation

No Name

state	Vector		Inserter		Ligation High ver.2
experiment	name	xx µg/mL	name	xx µg/mL	xx µg/mL
NC	name	xx µg/mL	name	xx µg/mL	xx µg/mL

Colony PCR

No name

Sample	base pair
9/8 Plac-spinach-DT-(1)	659

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5m	30s	30s	40s	30cycles

Sample	base pair
9/8 Plac-spinach-DT-(2)	659

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5m	30s	30s	40s	30cycles

Sample	base pair
9/8 Pcon-pt181atte-DT-(1)	584

PreDenature	Denature	Annealing	Extension	cycle
94°C	94°C	55°C	68°C	--
5m	30s	30s	40s	30cycles

Gel Extraction

じっけんしゃ

Lane	DNA	Enzyme
1	100bpladder	--
2	Spinach-DT	X+P
3	Spinach-DT	X+P
5	RBS-lacZα-DT	X+P
6	RBS-lacZα-DT	X+P

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

Name	quantity	concentration[µg/mL]	260/280	260/230
（´ω｀）
（ΦωΦ^）

Ligation

じっけんしゃ

state	Vector		Inserter		Ligation High ver.2
experiment	name	xx µg/mL	name	xx µg/mL	xx µg/mL
NC	name	xx µg/mL	name	xx µg/mL	xx µg/mL

PCR

Tatsui and Yoshida

Kai(100pg)	10x buffer KOD plus ver2	2M dNTP	MgSO4	Primer Mix(F&R)	Kod-plus ver2	MilliQ	total
0.5	2.5	2.5	1.5	1.5	0.5	16.0	25.0

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2m	10s	30s	2m	30cycles

Kai(400pg)	10x buffer KOD plus ver2	2M dNTP	MgSO4	Primer Mix(F&R)	Kod-plus ver2	MilliQ	total
2.0	2.5	2.5	1.5	1.5	0.5	14.5	25.0

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2m	10s	30s	2m	30cycles

Kai(800pg)	10x buffer KOD plus ver2	2M dNTP	MgSO4	Primer Mix(F&R)	Kod-plus ver2	MilliQ	total
4.0	2.5	2.5	1.5	1.5	0.5	12.5	25.0

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2m	10s	30s	2m	30cycles

Plac(100pg)	10x buffer KOD plus ver2	2M dNTP	MgSO4	Primer Mix(F&R)	Kod-plus ver2	MilliQ	total
0.3	2.5	2.5	1.5	1.5	0.5	16.2	25.0

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2m	10s	30s	2m	30cycles

Plac(200pg)	10x buffer KOD plus ver2	2M dNTP	MgSO4	Primer Mix(F&R)	Kod-plus ver2	MilliQ	total
0.6	2.5	2.5	1.5	1.5	0.5	15.9	25.0

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2m	10s	30s	2m	30cycles

Plac(400pg)	10x buffer KOD plus ver2	2M dNTP	MgSO4	Primer Mix(F&R)	Kod-plus ver2	MilliQ	total
1.2	2.5	2.5	1.5	1.5	0.5	15.3	25.0

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2m	10s	30s	2m	30cycles

Plac(800pg)	10x buffer KOD plus ver2	2M dNTP	MgSO4	Primer Mix(F&R)	Kod-plus ver2	MilliQ	total
2.4	2.5	2.5	1.5	1.5	0.5	14.1	25.0

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2m	10s	30s	2m	30cycles

Plac(nega)	10x buffer KOD plus ver2	2M dNTP	MgSO4	Primer Mix(F&R)	Kod-plus ver2	MilliQ	total
0.0	2.5	2.5	1.5	1.5	0.5	16.5	25.0

PreDenature	Denature	Annealing	Extension	cycle
94°C	98°C	57°C	68°C	--
2m	10s	30s	2m	30cycles

@@ Line 197: / Line 197: @@
 !Kai(100pg)||10x buffer KOD plus ver2||2M dNTP||MgSO4||Primer Mix(F&R)||Kod-plus ver2||MilliQ||total
 |-
-|0.5||2.5||2.5||1.5||1.5||0.5||16||25
+|0.5||2.5||2.5||1.5||1.5||0.5||16.0||25.0
+|}
+{| class="wikitable"
+!PreDenature||Denature||Annealing||Extension||cycle
+|-
+|94&deg;C||98&deg;C||57&deg;C||68&deg;C||--
+|-
+|2m||10s||30s||2m||30cycles
+|}
+</div>
+<!-- ここまでをコピーしてね -->
+<!-- ここから -->
+<div class="experiment">
+{| class="wikitable"
+!Kai(400pg)||10x buffer KOD plus ver2||2M dNTP||MgSO4||Primer Mix(F&R)||Kod-plus ver2||MilliQ||total
+|-
+|2.0||2.5||2.5||1.5||1.5||0.5||14.5||25.0
+|}
+{| class="wikitable"
+!PreDenature||Denature||Annealing||Extension||cycle
+|-
+|94&deg;C||98&deg;C||57&deg;C||68&deg;C||--
+|-
+|2m||10s||30s||2m||30cycles
+|}
+</div>
+<!-- ここまでをコピーしてね -->
+<!-- ここから -->
+<div class="experiment">
+{| class="wikitable"
+!Kai(800pg)||10x buffer KOD plus ver2||2M dNTP||MgSO4||Primer Mix(F&R)||Kod-plus ver2||MilliQ||total
+|-
+|4.0||2.5||2.5||1.5||1.5||0.5||12.5||25.0
+|}
+{| class="wikitable"
+!PreDenature||Denature||Annealing||Extension||cycle
+|-
+|94&deg;C||98&deg;C||57&deg;C||68&deg;C||--
+|-
+|2m||10s||30s||2m||30cycles
+|}
+</div>
+<!-- ここまでをコピーしてね -->
+<!-- ここから -->
+<div class="experiment">
+{| class="wikitable"
+!Plac(100pg)||10x buffer KOD plus ver2||2M dNTP||MgSO4||Primer Mix(F&R)||Kod-plus ver2||MilliQ||total
+|-
+|0.3||2.5||2.5||1.5||1.5||0.5||16.2||25.0
+|}
+{| class="wikitable"
+!PreDenature||Denature||Annealing||Extension||cycle
+|-
+|94&deg;C||98&deg;C||57&deg;C||68&deg;C||--
+|-
+|2m||10s||30s||2m||30cycles
+|}
+</div>
+<!-- ここまでをコピーしてね -->
+<!-- ここから -->
+<div class="experiment">
+{| class="wikitable"
+!Plac(200pg)||10x buffer KOD plus ver2||2M dNTP||MgSO4||Primer Mix(F&R)||Kod-plus ver2||MilliQ||total
+|-
+|0.6||2.5||2.5||1.5||1.5||0.5||15.9||25.0
+|}
+{| class="wikitable"
+!PreDenature||Denature||Annealing||Extension||cycle
+|-
+|94&deg;C||98&deg;C||57&deg;C||68&deg;C||--
+|-
+|2m||10s||30s||2m||30cycles
+|}
+</div>
+<!-- ここまでをコピーしてね -->
+<!-- ここから -->
+<div class="experiment">
+{| class="wikitable"
+!Plac(400pg)||10x buffer KOD plus ver2||2M dNTP||MgSO4||Primer Mix(F&R)||Kod-plus ver2||MilliQ||total
+|-
+|1.2||2.5||2.5||1.5||1.5||0.5||15.3||25.0
+|}
+{| class="wikitable"
+!PreDenature||Denature||Annealing||Extension||cycle
+|-
+|94&deg;C||98&deg;C||57&deg;C||68&deg;C||--
+|-
+|2m||10s||30s||2m||30cycles
+|}
+</div>
+<!-- ここまでをコピーしてね -->
+<!-- ここから -->
+<div class="experiment">
+{| class="wikitable"
+!Plac(800pg)||10x buffer KOD plus ver2||2M dNTP||MgSO4||Primer Mix(F&R)||Kod-plus ver2||MilliQ||total
+|-
+|2.4||2.5||2.5||1.5||1.5||0.5||14.1||25.0
+|}
+{| class="wikitable"
+!PreDenature||Denature||Annealing||Extension||cycle
+|-
+|94&deg;C||98&deg;C||57&deg;C||68&deg;C||--
+|-
+|2m||10s||30s||2m||30cycles
+|}
+</div>
+<!-- ここまでをコピーしてね -->
+<!-- ここから -->
+<div class="experiment">
+{| class="wikitable"
+!Plac(nega)||10x buffer KOD plus ver2||2M dNTP||MgSO4||Primer Mix(F&R)||Kod-plus ver2||MilliQ||total
+|-
+|0.0||2.5||2.5||1.5||1.5||0.5||16.5||25.0
 |}
 {| class="wikitable"

Template:Kyoto/Notebook/Sep 9

From 2013.igem.org

Revision as of 04:49, 24 September 2013

Contents

Sep 9

PCR

Miniprep

Restriction Enzyme Digestion

Transformation

Ligation

Colony PCR

Gel Extraction

Ligation

PCR