Template:Kyoto/Notebook/Aug 23

From 2013.igem.org

(Difference between revisions)
(→Electrophoresis)
(→Gel Extraction)
 
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Line 40: Line 40:
|17||Fusion1 attenuator||--||--
|17||Fusion1 attenuator||--||--
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:Igku_Aug23electrophoresis.jpg]]<br>
{| class="wikitable"
{| class="wikitable"
!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
Line 78: Line 78:
|17||tetR aptamer 12_P||--||--
|17||tetR aptamer 12_P||--||--
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:Igku_Aug23electrophoresis2.jpg]]<br>
<span class="author">Kojima and Honda</span>
<span class="author">Kojima and Honda</span>
{| class="wikitable"
{| class="wikitable"
Line 117: Line 117:
|17||tetR aptamer 12_1R||--||--
|17||tetR aptamer 12_1R||--||--
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:Igku Aug23 Electrophoresis(N2).jpg]]<br>
</div>
</div>
===Restriction Enzyme Digestion===
===Restriction Enzyme Digestion===
 +
<div class="experiment">
 +
<span class="author">Nakamoto and Kojima</span>
 +
{| class="wikitable"
 +
! ||Fusion6 antisense (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
 +
|-
 +
|2 cuts||3||0.5||0.5||3||0.3||22.7||30
 +
|-
 +
|1 cut||0.6||0.1||0||1||0.1||8.2||10
 +
|-
 +
|1 cut||0.6||0||0.1||1||0.1||8.2||10
 +
|-
 +
|NC||0.6||0||0||1||0.1||8.3||10
 +
|}
 +
{| class="wikitable"
 +
! ||Fusion6 antisense (1)||XbaI||PstI||buffer D||BSA||MilliQ||total
 +
|-
 +
|2 cuts||3||0.5||0.5||3||0.3||22.7||30
 +
|-
 +
|1 cut||0.6||0.1||0||1||0.1||8.2||10
 +
|-
 +
|1 cut||0.6||0||0.1||1||0.1||8.2||10
 +
|-
 +
|NC||0.6||0||0||1||0.1||8.3||10
 +
|}
 +
{| class="wikitable"
 +
! ||Fusion1 antisense (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
 +
|-
 +
|2 cuts||3.3||0.5||0.5||3||0.3||22.4||30
 +
|-
 +
|1 cut||0.7||0.1||0||1||0.1||8.1||10
 +
|-
 +
|1 cut||0.7||0||0.1||1||0.1||8.1||10
 +
|-
 +
|NC||0.7||0||0||1||0.1||8.2||10
 +
|}
 +
{| class="wikitable"
 +
! ||pT181 antisense (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
 +
|-
 +
|2 cuts||5.6||0.5||0.5||3||0.3||20.1||30
 +
|-
 +
|1 cut||1.1||0.1||0||1||0.1||7.7||10
 +
|-
 +
|1 cut||1.1||0||0.1||1||0.1||7.7||10
 +
|-
 +
|NC||1.1||0||0||1||0.1||7.8||10
 +
|}
 +
{| class="wikitable"
 +
! ||pT181 antisense (1)||XbaI||PstI||buffer D||BSA||MilliQ||total
 +
|-
 +
|2 cuts||5.6||0.5||0.5||3||0.3||20.1||30
 +
|-
 +
|1 cut||1.1||0.1||0||1||0.1||7.7||10
 +
|-
 +
|1 cut||1.1||0||0.1||1||0.1||7.7||10
 +
|-
 +
|NC||1.1||0||0||1||0.1||7.8||10
 +
|}
 +
*incubate at 37&deg;C from 17:29.
 +
</div>
===Electrophoresis===
===Electrophoresis===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp||--||--
 +
|-
 +
|2||pT181 attenuator||EcoRI||SpeI
 +
|-
 +
|3||pT181 attenuator||XbaI||PstI
 +
|-
 +
|4||Fusion3m2 attenuator||EcoRI||SpeI
 +
|-
 +
|5||Fusion1 attenuator||XbaI||PstI
 +
|-
 +
|6||Fusion1 attenuator||EcoRI||SpeI
 +
|-
 +
|7||Fusion3m2 attenuator||XbaI||PstI
 +
|-
 +
|8||pT181 antisense||EcoRI||SpeI
 +
|-
 +
|9||pT181 antisense||XbaI||PstI
 +
|-
 +
|10||Fusion1 antisense||EcoRI||SpeI
 +
|-
 +
|11||Fusion1 antisense||XbaI||PstI
 +
|-
 +
|12||Fusion6 antisense||EcoRI||SpeI
 +
|-
 +
|13||Fusion6 antisense||XbaI||PstI
 +
|-
 +
|14||tetR aptamer 12_1M||EcoRI||SpeI
 +
|-
 +
|15||tetR aptamer 12_P||EcoRI||SpeI
 +
|-
 +
|16||tetR aptamer 12_1R||EcoRI||SpeI
 +
|-
 +
|17||100bp||--||--
 +
|}
 +
[[File:Igku Aug23 Electrophoresis (N3).jpg]]<br>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp||--||--
 +
|-
 +
|2||Spinach||EcoRI||SpeI
 +
|-
 +
|3||J23100||SpeI||PstI
 +
|-
 +
|4||Plac||SpeI||PstI
 +
|-
 +
|5||100bp||--||--
 +
|-
 +
|6||J23100||SpeI||PstI
 +
|}
 +
[[File:Igku Aug23 Electrophoresis(N3)-2.jpg]]<br>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp||--||--
 +
|-
 +
|2||Fusion6 antisense||EcoRI||SpeI
 +
|-
 +
|3||Fusion6 antisense||EcoRI||--
 +
|-
 +
|4||Fusion6 antisense||--||SpeI
 +
|-
 +
|5||Fusion6 antisense||--||--
 +
|-
 +
|6||pT181 antisense||XbaI||PstI
 +
|-
 +
|8||100bp||--||--
 +
|-
 +
|9||100bp||--||--
 +
|-
 +
|10||Fusion6 antisense||XbaI||PstI
 +
|-
 +
|11||Fusion6 antisense||XbaI||--
 +
|-
 +
|12||Fusion6 antisense||--||PstI
 +
|-
 +
|13||Fusion6 antisense||--||--
 +
|-
 +
|14||pT181 antisense||XbaI||--
 +
|-
 +
|16||100bp||--||--
 +
|}
 +
[[File:Igku Aug23electrophoresis4.jpg]]<br>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp||--||--
 +
|-
 +
|2||Fusion1 antisense||EcoRI||SpeI
 +
|-
 +
|3||Fusion1 antisense||EcoRI||--
 +
|-
 +
|4||Fusion1 antisense||--||SpeI
 +
|-
 +
|5||Fusion1 antisense||--||--
 +
|-
 +
|6||pT181 antisense||--||PstI
 +
|-
 +
|8||100bp||--||--
 +
|-
 +
|9||100bp||--||--
 +
|-
 +
|10||pT181 antisense||EcoRI||SpeI
 +
|-
 +
|11||pT181 antisense||EcoRI||--
 +
|-
 +
|12||pT181 antisense||--||SpeI
 +
|-
 +
|13||pT181 antisense||--||--
 +
|-
 +
|14||pT181 antisense||--||--
 +
|-
 +
|16||100bp||--||--
 +
|}
 +
[[File:Igku Aug23electrophoresis5.jpg]]<br>
 +
 +
</div>
 +
===Gel Extraction===
===Gel Extraction===
 +
<div class="experiment">
 +
<span class="author">Nakamoto, Ashida, Kojima, Okazaki & Matsumoto</span>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|2||rowspan=2|pT181 attenuator||rowspan=2|XbaI & PstI
 +
|-
 +
|3
 +
|-
 +
|5||rowspan=2|Fusion3m2 attenuator||rowspan=2|EcoRI & SpeI
 +
|-
 +
|6
 +
|-
 +
|8||rowspan=2|Fusion1 attenuator||rowspan=2|XbaI & PstI
 +
|-
 +
|9
 +
|-
 +
|11||rowspan=2|Fusion1 attenuator||rowspan=2|EcoRI & SpeI
 +
|-
 +
|12
 +
|}
 +
[[File:Igku Aug23Gel Extraction.jpg]]<br>
 +
[[File:Igku Aug23 Gel Extraction(N6)-1.2.jpg]]<br>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|2||rowspan=2|Fusion3m2 attenuator||rowspan=2|XbaI & PstI
 +
|-
 +
|3
 +
|-
 +
|5||rowspan=2|pT181 antisense||rowspan=2|EcoRI & SpeI
 +
|-
 +
|6
 +
|-
 +
|8||rowspan=2|pT181 antisense||rowspan=2|XbaI & PstI
 +
|-
 +
|9
 +
|-
 +
|11||rowspan=2|Fusion1 antisense||rowspan=2|XbaI & PstI
 +
|-
 +
|12
 +
|}
 +
[[File:Igku Aug23 Gel Extraction(N7)-2.jpg]]<br>
 +
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|2||rowspan=2|Fusion6 attenuator||rowspan=2|XbaI & PstI
 +
|-
 +
|3
 +
|-
 +
|5||rowspan=2|aptamer 12_1M||rowspan=2|EcoRI & SpeI
 +
|-
 +
|6
 +
|}
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|2||rowspan=2|aptamer 12_P||rowspan=2|EcoRI & SpeI
 +
|-
 +
|3
 +
|-
 +
|5||rowspan=2|aptamer 12_1R||rowspan=2|EcoRI & SpeI
 +
|-
 +
|6
 +
|}
 +
[[File:Igku Aug23 Gel Extraction(N6)-2.4.jpg]]<br>
 +
[[File:Igku Aug23 Gel Extraction(N6)-3-2.jpg]]<br>
 +
<span class="author">Ashida</span>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|2||rowspan=2|Fusion6 antisense||rowspan=2|EcoRI & SpeI
 +
|-
 +
|3
 +
|-
 +
|5||rowspan=2|Fusion6 attenuator||rowspan=2|XbaI & PstI
 +
|-
 +
|6
 +
|-
 +
|8||rowspan=2|Fusion1 antisense||rowspan=2|EcoRI & SpeI
 +
|-
 +
|9
 +
|-
 +
|11||rowspan=2|pT181 antisense||rowspan=2|EcoRI & SpeI
 +
|-
 +
|12
 +
|}
 +
[[File:Igku Aug23 Gel Extracton(N8)-1.jpg]]<br>
 +
[[File:Igku Aug23 Gel Extraction(N8)-1.2.jpg]]<br>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|3||rowspan=2|pT181 antisense||rowspan=2|XbaI & PstI
 +
|-
 +
|4
 +
|}
 +
[[File:Igku Aug23 Gel Extraction(N7)-1.jpg]]<br>
 +
[[File:Igku Aug23 Gel Extraction(N7)1 2.jpg]]<br>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|3||rowspan=2|Plac||rowspan=2|SpeI & PstI
 +
|-
 +
|4
 +
|}
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|3||J23100||SpeI & PstI
 +
|-
 +
|5||rowspan=2|Spinach||rowspan=2|EcoRI & SpeI
 +
|-
 +
|6
 +
|}
 +
[[File:Igku Aug23 Gel Extraction(N9)-1-1.jpg]]<br>
 +
[[File:Igku Aug23 Gel Extraction(N9)-1-2.jpg]]<br>
 +
 +
 +
</div>

Latest revision as of 16:08, 25 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----

Igku Aug23electrophoresis.jpg

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----

Igku Aug23electrophoresis2.jpg
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----

Igku Aug23 Electrophoresis(N2).jpg

Restriction Enzyme Digestion

Nakamoto and Kojima

Fusion6 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
Fusion6 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
Fusion1 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
pT181 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
pT181 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----

Igku Aug23 Electrophoresis (N3).jpg

LaneSampleEnzyme1Enzyme2
1100bp----
2SpinachEcoRISpeI
3J23100SpeIPstI
4PlacSpeIPstI
5100bp----
6J23100SpeIPstI

Igku Aug23 Electrophoresis(N3)-2.jpg

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----

Igku Aug23electrophoresis4.jpg

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----

Igku Aug23electrophoresis5.jpg

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

Igku Aug23Gel Extraction.jpg
Igku Aug23 Gel Extraction(N6)-1.2.jpg

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

Igku Aug23 Gel Extraction(N7)-2.jpg

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

Igku Aug23 Gel Extraction(N6)-2.4.jpg
Igku Aug23 Gel Extraction(N6)-3-2.jpg
Ashida

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

Igku Aug23 Gel Extracton(N8)-1.jpg
Igku Aug23 Gel Extraction(N8)-1.2.jpg

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

Igku Aug23 Gel Extraction(N7)-1.jpg
Igku Aug23 Gel Extraction(N7)1 2.jpg

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

Igku Aug23 Gel Extraction(N9)-1-1.jpg
Igku Aug23 Gel Extraction(N9)-1-2.jpg


Retrieved from "http://2013.igem.org/Template:Kyoto/Notebook/Aug_23"