Template:Kyoto/Notebook/Aug 23

From 2013.igem.org

(Difference between revisions)
(→Gel Extraction)
(→Restriction Enzyme Digestion)
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<span class="author">Nakamoto and Kojima</span>
<span class="author">Nakamoto and Kojima</span>
{| class="wikitable"
{| class="wikitable"
-
!Fusion6 antisense (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
+
!state||Fusion6 antisense (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
|-
|-
-
|3||0.5||0.5||3||0.3||22.7||30
+
|2 cuts||3||0.5||0.5||3||0.3||22.7||30
|-
|-
-
|0.6||0.1||0||1||0.1||8.2||10
+
|1 cut||0.6||0.1||0||1||0.1||8.2||10
|-
|-
-
|0.6||0||0.1||1||0.1||8.2||10
+
|1 cut||0.6||0||0.1||1||0.1||8.2||10
|-
|-
-
|0.6||0||0||1||0.1||8.3||10
+
|NC||0.6||0||0||1||0.1||8.3||10
|}
|}
{| class="wikitable"
{| class="wikitable"
-
!Fusion6 antisense (1)||XbaI||PstI||buffer D||BSA||MilliQ||total
+
!state||Fusion6 antisense (1)||XbaI||PstI||buffer D||BSA||MilliQ||total
|-
|-
-
|3||0.5||0.5||3||0.3||22.7||30
+
|2 cuts||3||0.5||0.5||3||0.3||22.7||30
|-
|-
-
|0.6||0.1||0||1||0.1||8.2||10
+
|1 cut||0.6||0.1||0||1||0.1||8.2||10
|-
|-
-
|0.6||0||0.1||1||0.1||8.2||10
+
|1 cut||0.6||0||0.1||1||0.1||8.2||10
|-
|-
-
|0.6||0||0||1||0.1||8.3||10
+
|NC||0.6||0||0||1||0.1||8.3||10
|}
|}
{| class="wikitable"
{| class="wikitable"
-
!Fusion1 antisense (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
+
!state||Fusion1 antisense (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
|-
|-
-
|3.3||0.5||0.5||3||0.3||22.4||30
+
|2 cuts||3.3||0.5||0.5||3||0.3||22.4||30
|-
|-
-
|0.7||0.1||0||1||0.1||8.1||10
+
|1 cut||0.7||0.1||0||1||0.1||8.1||10
|-
|-
-
|0.7||0||0.1||1||0.1||8.1||10
+
|1 cut||0.7||0||0.1||1||0.1||8.1||10
|-
|-
-
|0.7||0||0||1||0.1||8.2||10
+
|NC||0.7||0||0||1||0.1||8.2||10
|}
|}
{| class="wikitable"
{| class="wikitable"
-
!pT181 antisense (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
+
!state||pT181 antisense (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
|-
|-
-
|5.6||0.5||0.5||3||0.3||20.1||30
+
|2 cuts||5.6||0.5||0.5||3||0.3||20.1||30
|-
|-
-
|1.1||0.1||0||1||0.1||7.7||10
+
|1 cut||1.1||0.1||0||1||0.1||7.7||10
|-
|-
-
|1.1||0||0.1||1||0.1||7.7||10
+
|1 cut||1.1||0||0.1||1||0.1||7.7||10
|-
|-
-
|1.1||0||0||1||0.1||7.8||10
+
|NC||1.1||0||0||1||0.1||7.8||10
|}
|}
{| class="wikitable"
{| class="wikitable"
-
!pT181 antisense (1)||XbaI||PstI||buffer D||BSA||MilliQ||total
+
!state||pT181 antisense (1)||XbaI||PstI||buffer D||BSA||MilliQ||total
|-
|-
-
|5.6||0.5||0.5||3||0.3||20.1||30
+
|2 cuts||5.6||0.5||0.5||3||0.3||20.1||30
|-
|-
-
|1.1||0.1||0||1||0.1||7.7||10
+
|1 cut||1.1||0.1||0||1||0.1||7.7||10
|-
|-
-
|1.1||0||0.1||1||0.1||7.7||10
+
|1 cut||1.1||0||0.1||1||0.1||7.7||10
|-
|-
-
|1.1||0||0||1||0.1||7.8||10
+
|NC||1.1||0||0||1||0.1||7.8||10
|}
|}
*incubate at 37&deg;C from 17:29.
*incubate at 37&deg;C from 17:29.

Revision as of 22:39, 20 September 2013

Contents

Aug 23

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1pT181 antisenseEcoRISpeI
2pT181 antisenseEcoRI--
3pT181 antisense--SpeI
4pT181 antisense----
5pT181 antisenseXbaIPstI
6pT181 antisenseXbaI--
7pT181 antisense--PstI
8pT181 antisense----
9100bp ladder----
10Fusion1 attenuatorEcoRISpeI
11Fusion1 attenuatorEcoRI--
12Fusion1 attenuator--SpeI
13Fusion1 attenuator----
14Fusion1 attenuatorXbaIPstI
15Fusion1 attenuatorXbaI--
16Fusion1 attenuator--PstI
17Fusion1 attenuator----

File:Igku xxxxxx.xxx

LaneSampleEnzyme1Enzyme2
1Fusion3m2 attenuatorEcoRISpeI
2Fusion3m2 attenuatorEcoRI--
3Fusion3m2 attenuator--SpeI
4Fusion3m2 attenuator----
5Fusion3m2 attenuatorXbaIPstI
6Fusion3m2 attenuatorXbaI--
7Fusion3m2 attenuator--PstI
8Fusion3m2 attenuator----
9100bp ladder----
10tetR aptamer 12_1MEcoRISpeI
11tetR aptamer 12_1MEcoRI--
12tetR aptamer 12_1M--SpeI
13tetR aptamer 12_1M----
14tetR aptamer 12_PEcoRISpeI
15tetR aptamer 12_PEcoRI--
16tetR aptamer 12_P--SpeI
17tetR aptamer 12_P----

File:Igku xxxxxx.xxx
Kojima and Honda

LaneSampleEnzyme1Enzyme2
1PlacSpeIPstI
2PlacSpeI--
3Plac--PstI
4Plac----
5SpinachEcoRISpeI
6SpinachEcoRI--
7Spinach--SpeI
8Spinach----
9100bp ladder----
10J23100SpeIPstI
11J23100SpeI--
12J23100--PstI
13J23100----
14tetR aptamer 12_1REcoRISpeI
15tetR aptamer 12_1REcoRI--
16tetR aptamer 12_1R--SpeI
17tetR aptamer 12_1R----

File:Igku xxxxxx.xxx

Restriction Enzyme Digestion

Nakamoto and Kojima

stateFusion6 antisense (1)EcoRISpeIbuffer BBSAMilliQtotal
2 cuts30.50.530.322.730
1 cut0.60.1010.18.210
1 cut0.600.110.18.210
NC0.60010.18.310
stateFusion6 antisense (1)XbaIPstIbuffer DBSAMilliQtotal
2 cuts30.50.530.322.730
1 cut0.60.1010.18.210
1 cut0.600.110.18.210
NC0.60010.18.310
stateFusion1 antisense (1)EcoRISpeIbuffer BBSAMilliQtotal
2 cuts3.30.50.530.322.430
1 cut0.70.1010.18.110
1 cut0.700.110.18.110
NC0.70010.18.210
statepT181 antisense (1)EcoRISpeIbuffer BBSAMilliQtotal
2 cuts5.60.50.530.320.130
1 cut1.10.1010.17.710
1 cut1.100.110.17.710
NC1.10010.17.810
statepT181 antisense (1)XbaIPstIbuffer DBSAMilliQtotal
2 cuts5.60.50.530.320.130
1 cut1.10.1010.17.710
1 cut1.100.110.17.710
NC1.10010.17.810
  • incubate at 37°C from 17:29.

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp----
2pT181 attenuatorEcoRISpeI
3pT181 attenuatorXbaIPstI
4Fusion3m2 attenuatorEcoRISpeI
5Fusion1 attenuatorXbaIPstI
6Fusion1 attenuatorEcoRISpeI
7Fusion3m2 attenuatorXbaIPstI
8pT181 antisenseEcoRISpeI
9pT181 antisenseXbaIPstI
10Fusion1 antisenseEcoRISpeI
11Fusion1 antisenseXbaIPstI
12Fusion6 antisenseEcoRISpeI
13Fusion6 antisenseXbaIPstI
14tetR aptamer 12_1MEcoRISpeI
15tetR aptamer 12_PEcoRISpeI
16tetR aptamer 12_1REcoRISpeI
17100bp----

File:Igku xxxxxx.xxx

LaneSampleEnzyme1Enzyme2
1100bp----
2SpinachEcoRISpeI
3J23100SpeIPstI
4PlacSpeIPstI
5100bp----

File:Igku xxxxxx.xxx

LaneSampleEnzyme1Enzyme2
1100bp----
2Fusion6 antisenseEcoRISpeI
3Fusion6 antisenseEcoRI--
4Fusion6 antisense--SpeI
5Fusion6 antisense----
6pT181 antisenseXbaIPstI
8100bp----
9100bp----
10Fusion6 antisenseXbaIPstI
11Fusion6 antisenseXbaI--
12Fusion6 antisense--PstI
13Fusion6 antisense----
14pT181 antisenseXbaI--
16100bp----

File:Igku xxxxxx.xxx

LaneSampleEnzyme1Enzyme2
1100bp----
2Fusion1 antisenseEcoRISpeI
3Fusion1 antisenseEcoRI--
4Fusion1 antisense--SpeI
5Fusion1 antisense----
6pT181 antisense--PstI
8100bp----
9100bp----
10pT181 antisenseEcoRISpeI
11pT181 antisenseEcoRI--
12pT181 antisense--SpeI
13pT181 antisense----
14pT181 antisense----
16100bp----

File:Igku xxxxxx.xxx

Gel Extraction

Nakamoto, Ashida, Kojima, Okazaki & Matsumoto

LaneDNAEnzyme
1100bp ladder--
2pT181 attenuatorXbaI & PstI
3
5Fusion3m2 attenuatorEcoRI & SpeI
6
8Fusion1 attenuatorXbaI & PstI
9
11Fusion1 attenuatorEcoRI & SpeI
12

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

LaneDNAEnzyme
1100bp ladder--
2Fusion3m2 attenuatorXbaI & PstI
3
5pT181 antisenseEcoRI & SpeI
6
8pT181 antisenseXbaI & PstI
9
11Fusion1 antisenseXbaI & PstI
12

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

LaneDNAEnzyme
1100bp ladder--
2Fusion6 attenuatorXbaI & PstI
3
5tetR aptamer 12_1MEcoRI & SpeI
6
LaneDNAEnzyme
1100bp ladder--
2tetR aptamer 12_PEcoRI & SpeI
3
5tetR aptamer 12_1REcoRI & SpeI
6

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

LaneDNAEnzyme
1100bp ladder--
2Fusion6 antisenseEcoRI & SpeI
3
5Fusion6 attenuatorXbaI & PstI
6
8Fusion1 antisenseEcoRI & SpeI
9
11pT181 antisenseEcoRI & SpeI
12

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

LaneDNAEnzyme
1100bp ladder--
3pT181 antisenseXbaI & PstI
4

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

LaneDNAEnzyme
1100bp ladder--
3PlacSpeI & PstI
4
LaneDNAEnzyme
1100bp ladder--
3J23100SpeI & PstI
5SpinachEcoRI & SpeI
6

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

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