Template:Kyoto/Notebook/Sep 9

From 2013.igem.org

(Difference between revisions)
(→Colony PCR)
(→Liquid Culture)
Line 334: Line 334:
|-
|-
|2m||10s||30s||2m||30cycles
|2m||10s||30s||2m||30cycles
 +
|}
 +
</div>
 +
<!-- ここまでをコピーしてね -->
 +
===Liquid Culture===
 +
<!-- ここから -->
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|||
 +
|-
 +
|||
 +
|-
 +
|||
 +
|-
 +
|||
 +
|}
 +
</div>
 +
<!-- ここまでをコピーしてね -->
 +
===Restriction Enzyme Digestion===
 +
<!-- こっから -->
 +
<div class="experiment">
 +
<span class="author">じっけんしゃ</span>
 +
{| class="wikitable"
 +
! ||DNA||restricion enzyme||restriction enzyme||Buffer||BSA||MilliQ||total
 +
|-
 +
|2 cuts||（°°） &micro;L|| &micro;L|| &micro;L|| &micro;L|| &micro;L|| &micro;L|| &micro;L
 +
|-
 +
|NC|| &micro;L|| &micro;L|| &micro;L|| &micro;L|| &micro;L|| &micro;L|| &micro;L
 +
|}
 +
</div>
 +
<!-- ここまでをコピーしてね -->
 +
===PCR===
 +
<!-- ここから -->
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!genome DNA||KOD plus||10x buffer||dNTP||MgSO4||SasA_fwd primer||SasA_rev primer||MilliQ||total
 +
|-
 +
| ||0.5||2.5||2.5||1.5||0.75||0.75|| ||25
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|　&deg;C||　&deg;C||　&deg;C||　&deg;C||--
 +
|-
 +
|（◎Д◎）　||　||　||　||
|}
|}
</div>
</div>
Line 354: Line 402:
</div>
</div>
<!-- ここまでをコピーしてね -->
<!-- ここまでをコピーしてね -->
 +
===Gel Extraction===
===Gel Extraction===
<!-- こっから -->
<!-- こっから -->

Revision as of 08:24, 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

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

Fe(Ⅲ)

Shimizu & Hirano

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

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

Gel Extraction

じっけんしゃ

LaneDNAEnzyme
1100bp ladder--
2Spinach-DTXbaI+PstI
3
5RBS-lacZα-DTXbaI+PstI
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

Liquid Culture

No name

Samplemedium

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

PCR

No name

genome DNAKOD plus10x bufferdNTPMgSO4SasA_fwd primerSasA_rev primerMilliQtotal
0.52.52.51.50.750.75 25
PreDenatureDenatureAnnealingExtensioncycle
　°C　°C　°C　°C--
（◎Д◎）　　　　

Liquid Culture

No name

Samplemedium
9/2 Spinach-DTPlusgrow medium(+CP)
8/29 Plux-RBS-GFP-DTPlusgrow medium
9/6 master plate Plusgrow medium(+Amp)
3 Pcon-PT181 attenuatorPlusgrow medium

Gel Extraction

No name

LaneDNAEnzyme
11kbp ladder--
3lysis1XbaI+PstI
4
5
6

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

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

No name

LaneDNAEnzyme
11kbp ladder--
3lysis3XbaI+PstI
4
5
6

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

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

Electrophoresis

Yoshida

LaneSampleEnzyme1Enzyme2
11kb ladder----
2Plac 100pg----
3Plac 200pg----
4Plac 400pg----
5Plac 800pg----
6NC----
71kb ladder----

File:Igku xxxxxx.xxx

Retrieved from "http://2013.igem.org/Template:Kyoto/Notebook/Sep_9"