Template:Kyoto/Notebook/Sep 16

From 2013.igem.org

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!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2
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|experiment||name||xx µg/mL||name||xx µg/mL||xx µg/mL
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|experiment||Pcon-pT181anti-spin-DT(X+P)||5.0µg/mL||pSB4K5(X+P)||18.1µg/mL||xx µg/mL
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|NC||name||xx µg/mL||name||xx µg/mL||xx µg/mL
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|experiment||pC181 attenuater(X+P)||8.2µg/mL||pSB1C3(X+P)||26.0µg/mL||xx µg/mL
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===PCR===
===PCR===
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Revision as of 06:37, 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
1100bp ladder
2anti
3anti
4anti
5tet aptamer-DT
6tet aptamer-DT
7tet aptamer-DT
8nega

File:Igku xxxxxx.xxx

Restriction Enzyme Digestion

じっけんしゃ

9/16 Pcon-RBS-tetR-DT(µL)E(µL)S(µL)X(µL)P(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts15.80011336.230
NC0.80000117.210
9/16 pSB4K5(µL)E(µL)S(µL)X(µL)P(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts13.50011338.530
NC0.70000117.310
8/22 pSB1C3(µL)E(µL)S(µL)X(µL)P(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts18.51001333.530
NC0.90000117.110
8/22 pSB1C3(µL)E(µL)S(µL)X(µL)P(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts18.51100333.530
NC0.90000117.110
8/21 pT181 attenuator2(µL)E(µL)S(µL)X(µL)P(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts7.411003314.630
NC0.40000117.610

Ligation

じっけんしゃ

stateVectorInserterLigation High ver.2
experimentPcon-pT181anti-spin-DT(X+P)5.0µg/mLpSB4K5(X+P)18.1µg/mLxx µg/mL
experimentpC181 attenuater(X+P)8.2µg/mLpSB1C3(X+P)26.0µg/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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