Template:Kyoto/Notebook/Sep 16

From 2013.igem.org

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(→Miniprep)
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{| class="wikitable"
{| class="wikitable"
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!genome DNA||KOD plus||10x buffer||dNTP||MgSO4||SasA_fwd primer||SasA_rev primer||MilliQ||total
+
{| class="wikitable"
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!9/6pT181 anti psB1C3(1)||KOD plus||10x buffer||dNTP||MgSO4||GG1_1C3_fwd primer||GG2_1C3_rev primer||MilliQ||total
|-
|-
-
| ||0.5||2.5||2.5||1.5||0.75||0.75|| ||25
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|1||0.5||2.5||2.5||1.5||0.75||0.75||15.5||25
|}
|}
{| class="wikitable"
{| class="wikitable"
!PreDenature||Denature||Annealing||Extension||cycle
!PreDenature||Denature||Annealing||Extension||cycle
|-
|-
-
|　&deg;C||　&deg;C||　&deg;C||　&deg;C||--
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|94&deg;C||98&deg;C||57&deg;C||68&deg;C||30cycles
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|}
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{| class="wikitable"
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!9/6 12-1R aptamer-DT||KOD plus||10x buffer||dNTP||MgSO4||GG1_1C3_fwd primer||GG1_1C3_rev primer||MilliQ||total
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|-
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|1||0.5||2.5||2.5||1.5||0.75||0.75||15.5||25
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|}
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{| class="wikitable"
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!PreDenature||Denature||Annealing||Extension||cycle
|-
|-
-
|（◎Д◎）　||　||　||　||
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|94&deg;C||98&deg;C||57&deg;C||68&deg;C||30cycles
|}
|}
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===Electrophoresis===
===Electrophoresis===
   
   

Revision as of 06:22, 24 September 2013

Contents

Sep 16

Liquid Culture

No name

Samplemedium
pSB1C3+P-KaiBC①Plusgrow medium(+CP)
pSB1C3+P-KaiBC②Plusgrow medium(+CP)

Colony PCR

No name

Samplebase pair
pSB1C3+P-KaiBC①283
pSB1C3+P-KaiBC②283
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
2min30s30s18s30cycles

Gel Extraction

じっけんしゃ

LaneDNAEnzyme
11kb ladder
2pT181 attenuatorX+P
3pT181 attenuatorX+P
4pT181 attenuatorX+P
6pSB1C3E+S
7pSB1C3E+S
8pSB1C3E+S
10pSB4K5X+P
11pSB4K5X+P
12pSB4K5X+P

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

Nameconcentration[µg/mL]260/280260/230
pT181 attenuator(X+P)8.21.620.11
pSB1C3(E+S)49.51.480.18
pSB4K5(X+P)18.11.730.57
DT(S+P)70.41.361.02
aptamer12-1R-DT(S+P)42.01.820.47
spinach-DT(S+P)36.81.790.61

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
Dx-lux1136.71.461.28
RBS-laczα-DT237.61.881.70
pSB4K5148.41.551.09
Pcon-KBS-tetR-DT126.61.731.36
9/11Plac1160.01.781.28
9/15Plac2144.01.731.81
9/14Plac332.51.512.33
Ptet-RBS-laczα-DT231.31.841.57

PCR

No name

9/6pT181 anti psB1C3(1)KOD plus10x bufferdNTPMgSO4GG1_1C3_fwd primerGG2_1C3_rev primerMilliQtotal
10.52.52.51.50.750.7515.525
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C30cycles
9/6 12-1R aptamer-DTKOD plus10x bufferdNTPMgSO4GG1_1C3_fwd primerGG1_1C3_rev primerMilliQtotal
10.52.52.51.50.750.7515.525
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C30cycles

Electrophoresis

じっけんしゃ

LaneSampleEnzyme1Enzyme2
（°Д°）

File:Igku xxxxxx.xxx

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

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

No name

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