Template:Kyoto/Notebook/Sep 7

From 2013.igem.org

(Difference between revisions)
(Ligation)
(Electrophoresis)
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</div>
</div>
===Electrophoresis===
===Electrophoresis===
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<div class="experiment">
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<span class="author">No name</span>
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{| class="wikitable"
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!Lane||Sample
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|-
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|1||100bp ladder
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|-
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|2||Plac-Spinach-DT -5
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|-
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|3||Plac-Spinach-DT -6
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|-
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|4||Plac-Spinach-DT -7
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|-
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|5||Plac-Spinach-DT -8
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|-
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|6||Plac-Spinach-DT -9
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|-
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|7||Plac-Spinach-DT -10
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|-
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|8||100bp ladder
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|-
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|1||100bp ladder
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|-
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|2||Plac-Spinach-DT -11
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|-
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|3||Plac-Spinach-DT -12
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|-
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|4||Pcon-antisense -5
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|-
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|5||Pcon-antisense -6
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|-
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|6||Pcon-antisense -7
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|-
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|7||Pcon-antisense -8
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|-
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|8||100bp ladder
 +
|}
 +
[[File:igku_xxxxxx.xxx]]<br>
 +
{| class="wikitable"
 +
!Lane||Sample
 +
|-
 +
|1||100bp ladder
 +
|-
 +
|2||Pcon-antisense -9
 +
|-
 +
|3||Pcon-antisense -10
 +
|-
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|4||Pcon-antisense -11
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|-
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|5||Pcon-antisense -12
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|-
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|6||Plac-attenuator -5
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|-
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|7||Plac-attenuator -6
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|-
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|8||100bp ladder
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|-
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|1||100bp ladder
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|-
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|2||Plac-attenuator -7
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|-
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|3||Plac-attenuator -8
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|-
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|4||Plac-attenuator -9
 +
|-
 +
|5||Plac-attenuator -10
 +
|-
 +
|6||Plac-attenuator -11
 +
|-
 +
|7||Plac-attenuator -12
 +
|-
 +
|8||100bp ladder
 +
|}
 +
[[File:igku_xxxxxx.xxx]]<br>
 +
{| class="wikitable"
 +
!Lane||Sample
 +
|-
 +
|1||100bp ladder
 +
|-
 +
|2||Plac-pT181 antisense
 +
|}
 +
[[File:igku_xxxxxx.xxx]]<br>
 +
</div>
 +
===Ligation===
===Ligation===
<div class="experiment">
<div class="experiment">

Revision as of 12:47, 25 September 2013

Contents

Sep 7

Igku Sep7electrophoresis1.jpg Igku Sep7electrophoresis2.jpg Igku Sep7electrophoresis3.jpg

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
RBS-lysis1-DT195.3
RBS-lysis3-DT255.5 1.91 2.04
Ptet-pT18183.6 1.85 1.48
Pcon-RBS-tetR-DT134.3 1.94 2.20
Ptet-RBS-lacZα-DT67.7 1.92 1.79
Pcon-pT181 attenuator232.3 1.95 1.90
pSB4K5237.0 1.61 1.50
pT181 antisense(pSB1C3)256.4 1.63 1.78
apt12_1R(pSB1C3)230.0 1.98 2.28

Electrophoresis

No name

LaneSample
1100bp ladder
2Plac-Spinach-DT -5
3Plac-Spinach-DT -6
4Plac-Spinach-DT -7
5Plac-Spinach-DT -8
6Plac-Spinach-DT -9
7Plac-Spinach-DT -10
8100bp ladder
1100bp ladder
2Plac-Spinach-DT -11
3Plac-Spinach-DT -12
4Pcon-antisense -5
5Pcon-antisense -6
6Pcon-antisense -7
7Pcon-antisense -8
8100bp ladder

File:Igku xxxxxx.xxx

LaneSample
1100bp ladder
2Pcon-antisense -9
3Pcon-antisense -10
4Pcon-antisense -11
5Pcon-antisense -12
6Plac-attenuator -5
7Plac-attenuator -6
8100bp ladder
1100bp ladder
2Plac-attenuator -7
3Plac-attenuator -8
4Plac-attenuator -9
5Plac-attenuator -10
6Plac-attenuator -11
7Plac-attenuator -12
8100bp ladder

File:Igku xxxxxx.xxx

LaneSample
1100bp ladder
2Plac-pT181 antisense

File:Igku xxxxxx.xxx

Ligation

No Name

stateVectorInserterLigation High ver.2
experiment9/6 Plux (SpeI&PstI)2 µL9/7 RBS-lysis2-DT (EcoRI&XbaI) µL3.5 µL
experiment9/6 Plac (SpeI&PstI) µL9/7 RBS-lysis2-DT (EcoRI&XbaI) µL3.5 µL
experiment9/6 Pcon-RBS-luxR-DT (EcoRI&XbaI)1.9 µL9/6 Plux-RBS-GFP-DT(SpeI&PstI)2.2 µL3.5 µL
experiment9/6 Pbed/araC (SpeI&PstI)1.6 µL9/5 RBS-luxI-DT (EcoRI&XbaI)16 µL3.5 µL
experiment9/6 Plac (SpeI&PstI)1.5 µL9/5 pT181 antisense (EcoRI&XbaI)2.0 µL3.5 µL
experiment9/6 Pcon (SpeI&PstI)3.0 µL9/5 RBS-luxI-DT (EcoRI&XbaI)10 µL3.5 µL

Colony PCR

No name

Samplebase pair
9/5 Plac-spinarch-DT-13~20
9/5 Plac-pT181 attenuator-13~20
PreDenatureDenatureAnnealingExtensioncycle
94 °C94 °C55 °C60 °C--
5min 30s 30s 42s 30cycle

Transformation

Tatsui

NameSampleCompetent CellsTotalPlate
Pcon-RBS-luxR-DT+Plux-RBS-GFP-DT2µL20µL22µL
Pbed/araC+RBS-luxI-DT2µL 20µL 22µL
Plac+pT181 antisence 2µL 20µL 22µL
Pcon+RBS-luxI-DT 2µL 20µL 22µL

Restriction Enzyme Digestion

No Name

DNAXbaIPstIBufferBSAMilliQtotal
9/7 RBS-lysis1-DT(XbaI & PstI)10 µL1 µL1 µL3 µL3 µL12 µL30 µL
NC0.5 µL0 µL0 µL1 µL1 µL7.5 µL10 µL


DNAXbaIPstIBufferBSAMilliQtotal
9/7 RBS-lysis3-DT (XbaI & PstI)7.8 µL1 µL1 µL3 µL3 µL14.2 µL30 µL
NC0.4 µL0 µL0 µL1 µL1 µL7.6 µL10 µL


DNASpeIPstIBufferBSAMilliQtotal
9/5 Plac (SpeI & PstI)6.0 µL1 µL1 µL3 µL3 µL16.0 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNAEcorISpeIBufferBSAMilliQtotal
9/7 pSB4K5 (EcoRI & SpeI)8.4 µL1 µL1 µL3 µL3 µL16.0 µL30 µL
NC0.4 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNArestricion enzymerestriction enzymeBufferBSAMilliQtotal
Pcon-pT181 attenuator (SpeI & PstI)8.6 µL1 µL1 µL3 µL3 µL13.4 µL30 µL
Pcon-pT181 attenuator (EcorI & SpeI)8.6 µL1 µL1 µL3 µL3 µL13.4 µL30 µL
NC0.4 µL0 µL0 µL1 µL1 µL7.6 µL10 µL


DNAXbaIPstIBufferBSAMilliQtotal
9/5 Spin-DT (XbaI & PstI)5.4 µL1 µL1 µL3 µL3 µL16.6 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNASpeIPstIBufferBSAMilliQtotal
Ptet-anticodone (SpeI & PstI)11 µL1 µL1 µL3 µL3 µL11 µL30 µL
NC0.5 µL0 µL0 µL1 µL1 µL7.5 µL10 µL


DNAXbaIPstIBufferBSAMilliQtotal
9/6 RBS-lacZα-DT (XbaI & PstI)11 µL1 µL1 µL3 µL3 µL11 µL30 µL
NC0.5 µL0 µL0 µL1 µL1 µL7.5 µL10 µL


DNAEcoRISpeIBufferBSAMilliQtotal
pSB1C3 (1) (EcoRI & SpeI)6.6 µL1 µL1 µL3 µL3 µL15.4 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNAXbaIPstIBufferBSAMilliQtotal
pSB1C3 (3) (XbaI & PstI)6.6 µL1 µL1 µL3 µL3 µL15.4 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL