Template:Kyoto/Notebook/Sep 9

From 2013.igem.org

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===Gel Extraction===
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<div class="experiment">
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<span class="author">じっけんしゃ</span>
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{| class="wikitable"
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!Lane||DNA||Enzyme
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|-
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|（＞ω・）b || ||
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[[File:igku_xxbeforexx.xxx]]<br>
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[[File:igku_xxafterxx.xxx]]<br>
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{| class="wikitable"
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!Name||concentration[&micro;g/mL]||260/280||260/230
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|（´ω｀）|| || ||
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|（ΦωΦ^）|| || ||
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<!-- ここまでをコピーしてね -->
===Miniprep===
===Miniprep===

Revision as of 06:49, 24 September 2013

Contents

Sep 9

PCR

Nakagawa

KaiA(plasmid)10x bufferdNTPsMgSO4Primer(661-RBS-KaiA-fwd)Primer(kaiA-662-rev-ver2)Kod-plus ver2MilliQtotal
1.02.52.51.50.750.750.515.525
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C58.5°C68°C--
2min10s30s30s30cycles
KaiB(plasmid)10x bufferdNTPsMgSO4Primer(661-RBS-KaiA-fwd-ver2)Primer(kaiB-663-rev-ver2)Kod-plus ver2MilliQtotal
1.02.52.51.50.750.750.515.525
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C58.5°C68°C--
2min10s30s30s30cycles
P-KaiBC(PCR products)10x bufferdNTPsMgSO4Primer(prefix-P-kaiBC-fwd-ver2)Primer(P-KaiBC-suffix-rev-verw)Kod-plus ver2MilliQtotal
0.72.52.51.50.750.750.515.825
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C58.5°C68°C--
2min10s30s30s30cycles
DT(plasmid)10x bufferdNTPsMgSO4Primer(664-DT-fwd-ver2)Primer(DT-suffix-660-rev-ver2)Kod-plus ver2MilliQtotal
1.02.52.51.50.750.750.515.525
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C58.5°C68°C--
2min10s30s30s30cycles

Gel Extraction

じっけんしゃ

LaneDNAEnzyme
（＞ω・）b

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

Nameconcentration[µg/mL]260/280260/230
（´ω｀）
（ΦωΦ^）

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
9/8 pSB1C383.21.450.76
9/8 RBS-lysis2-DT72.81.670.97
9/8 spinach-DT152.61.881.97
Nuclease-Free water-0.4-14.300.25

Restriction Enzyme Digestion

じっけんしゃ

DNArestricion enzymerestriction enzymeBufferBSAMilliQtotal
2 cuts（°°） µL µL µL µL µL µL µL
NC µL µL µL µL µL µL µL

Transformation

じっけんしゃ

NameSampleCompetent CellsTotalPlate
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

stateVectorInserterLigation High ver.2
experimentnamexx µg/mLnamexx µg/mLxx µg/mL
NCnamexx µg/mLnamexx µg/mLxx µg/mL

Colony PCR

No name

Samplebase pair
9/8 Plac-spinach-DT-(1)659
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s40s30cycles
Samplebase pair
9/8 Plac-spinach-DT-(2)659
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s40s30cycles
Samplebase pair
9/8 Pcon-pt181atte-DT-(1)584
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s40s30cycles

Gel Extraction

じっけんしゃ

LaneDNAEnzyme
1100bpladder--
2Spinach-DTX+P
3
5RBS-lacZα-DTX+P
6

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

Namequantityconcentration[µg/mL]260/280260/230
（´ω｀）
（ΦωΦ^）

Ligation

じっけんしゃ

stateVectorInserterLigation High ver.2
experimentnamexx µg/mLnamexx µg/mLxx µg/mL
NCnamexx µg/mLnamexx µg/mLxx µg/mL

PCR

Tatsui and Yoshida

Kai(100pg)10x buffer KOD plus ver22M dNTPMgSO4Primer Mix(F&R)Kod-plus ver2MilliQtotal
0.52.52.51.51.50.516.025.0
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2m10s30s2m30cycles
Kai(400pg)10x buffer KOD plus ver22M dNTPMgSO4Primer Mix(F&R)Kod-plus ver2MilliQtotal
2.02.52.51.51.50.514.525.0
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2m10s30s2m30cycles
Kai(800pg)10x buffer KOD plus ver22M dNTPMgSO4Primer Mix(F&R)Kod-plus ver2MilliQtotal
4.02.52.51.51.50.512.525.0
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2m10s30s2m30cycles
Plac(100pg)10x buffer KOD plus ver22M dNTPMgSO4Primer Mix(F&R)Kod-plus ver2MilliQtotal
0.32.52.51.51.50.516.225.0
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2m10s30s2m30cycles
Plac(200pg)10x buffer KOD plus ver22M dNTPMgSO4Primer Mix(F&R)Kod-plus ver2MilliQtotal
0.62.52.51.51.50.515.925.0
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2m10s30s2m30cycles
Plac(400pg)10x buffer KOD plus ver22M dNTPMgSO4Primer Mix(F&R)Kod-plus ver2MilliQtotal
1.22.52.51.51.50.515.325.0
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2m10s30s2m30cycles
Plac(800pg)10x buffer KOD plus ver22M dNTPMgSO4Primer Mix(F&R)Kod-plus ver2MilliQtotal
2.42.52.51.51.50.514.125.0
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2m10s30s2m30cycles
Plac(nega)10x buffer KOD plus ver22M dNTPMgSO4Primer Mix(F&R)Kod-plus ver2MilliQtotal
0.02.52.51.51.50.516.525.0
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2m10s30s2m30cycles
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