Template:Kyoto/Notebook/Sep 8

From 2013.igem.org

(Difference between revisions)
(→Restriction Enzyme Digestion)
(→Transformation)
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<span class="author">じっけんしゃ</span>
<span class="author">じっけんしゃ</span>
{| class="wikitable"
{| class="wikitable"
-
!Name||Sample||Competent Cells||Total||Plate
+
!Name||Sample(&micro;L)||Competent Cells(&micro;L)||Total(&micro;L)||Plate
|-
|-
-
|いでんし||なんちゃら&micro;L||ごにょごにょ&micro;L||ほにゃらら&micro;L||こーせーぶっしつ
+
|9/8 Pcont+aptamer 12-1R-DT||1||10||11||Amp
|-
|-
-
|いかどうよう|| || || ||
+
|9/8 Pcon+RBS-lacZα-DT||1||10||11||Amp
|-
|-
-
| || || || ||
+
|9/8 Pcon-pT181 attenuater+aptamer 12-1R-DT||1||10||11||Amp
 +
|-
 +
|9/8 Plac+RBS-lysis1-DT||2||20||22||CP
 +
|-
 +
|9/8 Plac+RBS-lysis2-DT||2||20||22||CP
 +
|-
 +
|9/8 Plac+aptamer 12-1R-DT||2||20||22||CP
 +
|-
 +
|9/8 Plac+spinach||2||20||22||CP
 +
|-
 +
|9/8 Plac+pT181 attenuater||2||20||22||CP
 +
|-
 +
|9/8 Plux+RBS-lysis1-DT||1||10||11||CP
 +
|-
 +
|9/8 Plux+RBS-lysis2-DT||1||10||11||CP
 +
|-
 +
|9/8 Ptet+RBS-lacZα-DT||1||10||11||CP
 +
|-
 +
|9/8 Ptet-pT181 anti+spinach-DT||1||10||11||CP
 +
|-
 +
|9/8 Pcon-pT181 attenuater+DT||1||10||11||CP
|}
|}
 +
===Electrophoresis===
===Electrophoresis===
   
   

Revision as of 01:19, 24 September 2013

Contents

Sep 8

Gel Extraction

No name

LaneDNAEnzyme
1100bp ladder
2RBS-lysis1-DTXbaI & PstI
3RBS-lysis1-DTXbaI & PstI
5RBS-lysis3-DTXbaI & PstI
6RBS-lysis3-DTXbaI & PstI

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

Nameconcentration[µg/mL]260/280260/230
RBS-lysis1-DT (XbaI & PstI)5.31.980.06
RBS-lysis3-DT (XbaI & PstI)5.31.980.06
LaneDNAEnzyme
1100bp ladder
3Pcon-pT181 attenuatorEcoRI & SpeI
4Pcon-pT181 attenuatorEcoRI & SpeI
6Spinach-DTXbaI & PstI
7Spinach-DTXbaI & PstI
9RBS-lacZα-DTXbaI & PstI
10RBS-lacZα-DTXbaI & PstI

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

Nameconcentration[µg/mL]260/280260/230
Pcon-pT181 attenuator (EcoRI & SpeI)5.21.820.36
Spinach-DT (XbaI & PstI)8.01.870.25
RBS-lacZα-DT (XbaI & PstI)10.31.710.54
LaneDNAEnzyme
11kbp ladder
2Ptet-pT181 antisenseEcoRI & PstI
3Ptet-pT181 antisenseEcoRI & PstI
5pSB4K5EcoRI & SpeI
6pSB4K5EcoRI & SpeI

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

Nameconcentration[µg/mL]260/280260/230
Ptet-pT181 antisense(EcoRI & PstI)30.61.841.16
pSB4K5 (EcoRI & SpeI)21.81.870.98
LaneDNAEnzyme
11kbp ladder
2Pcon-pT181 attenuatorSpeI & PstI
3Pcon-pT181 attenuatorSpeI & PstI
5PlacSpeI & PstI
6PlacSpeI & PstI
8pSB1C3EcoRI & SpeI
9pSB1C3EcoRI & SpeI
11pSB1C3XbaI & PstI
12pSB1C3XbaI & PstI

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

Nameconcentration[µg/mL]260/280260/230
Pcon-pT181 attenuator (SpeI & PstI)14.51.911.09
Plac (SpeI & PstI)11.61.741.19
pSB1C3 (EcoRI & SpeI)5.71.740.34
pSB1C3 (XbaI & PstI)5.62.000.37

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
Pcon-pT181 anti243.01.901.81
Pcon-Spinach-DT253.71.571.51

Restriction Enzyme Digestion

じっけんしゃ

9/8 Pcon-pT181 antiESXPBSABufferMilliQtotal
2 cuts8.2µL 0µL 1µL 0µL 1µL 3µL 3µL 13.8µL 30µL
NC 0.4µL 0µL 0µL 0µL 0µL 1µL 1µL 7.6µL 10µL
9/8 Pcon-spin-DTESXPBSABufferMilliQtotal
2 cuts7.9µL 1µL 1µL 0µL 0µL 3µL 3µL14.1µL 30µL
NC 0.4µL 0µL 0µL 0µL 0µL 1µL 1µL7.6µL 10µL
8/21 F1 atteESXPBSABufferMilliQtotal
2 cuts 6.5µL 0µL 0µL 1µL 1µL 3µL 3µL15.5µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 F3m2 atteESXPBSABufferMilliQtotal
2 cuts6.0µL0µL0µL1µL1µL3µL3µL16µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 F1 antiESXPBSABufferMilliQtotal
2 cuts6.7µL0µL0µL1µL1µL3µL3µL15.3µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 F6 antiESXPBSABufferMilliQtotal
2 cuts6.0µL0µL0µL1µL1µL3µL3µL16µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 tetR aptamer 12_1MESXPBSABufferMilliQtotal
2 cuts5.3µL1µL1µL0µL0µL3µL3µL16.7µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 tetR aptamer 12_PESXPBSABufferMilliQtotal
2 cuts13.3µL1µL1µL0µL0µL3µL3µL8.7µL30µL
NC0.7µL0µL0µL0µL0µL1µL1µL7.3µL10µL
9/4 spin-DTESXPBSABufferMilliQtotal
2 cuts5.4µL0µL0µL1µL1µL3µL3µL16.6µL30µL
NC0.3µL0µL0µL0µL0µL1µL1µL7.7µL10µL
9/7 RBS-lysis1-DTESXPBSABufferMilliQtotal
2 cuts10µL0µL0µL1µL1µL3µL3µL12µL30µL
NC0.5µL0µL0µL0µL0µL1µL1µL7.5µL10µL
9/6 RBS-lysis2-DTESXPBSABufferMilliQtotal
2 cuts13µL0µL0µL1µL1µL3µL3µL9µL30µL
NC0.7µL0µL0µL0µL0µL1µL1µL7.3µL10µL
9/7 RBS-lysis3-DTESXPBSABufferMilliQtotal
2 cuts10µL0µL0µL1µL1µL3µL3µL12µL30µL
NC0.5µL0µL0µL0µL0µL1µL1µL7.5µL10µL
8/20 J23100 RBS-lacZα-DT-(1)ESXPBSABufferMilliQtotal
2 cuts7µL0.5µL0.5µL0µL0µL3µL3µL16µL30µL
NC0.5µL0µL0µL0µL0µL1µL1µL7.5µL10µL
8/20 RBS-lacZα-DTESXPBSABufferMilliQtotal
2 cuts10µL0.5µL0.5µL0µL0µL3µL3µL13µL30µL
NC0.5µL0µL0µL0µL0µL1µL1µL7.5µL10µL

Transformation

じっけんしゃ

NameSample(µL)Competent Cells(µL)Total(µL)Plate
9/8 Pcont+aptamer 12-1R-DT11011Amp
9/8 Pcon+RBS-lacZα-DT11011Amp
9/8 Pcon-pT181 attenuater+aptamer 12-1R-DT11011Amp
9/8 Plac+RBS-lysis1-DT22022CP
9/8 Plac+RBS-lysis2-DT22022CP
9/8 Plac+aptamer 12-1R-DT22022CP
9/8 Plac+spinach22022CP
9/8 Plac+pT181 attenuater22022CP
9/8 Plux+RBS-lysis1-DT11011CP
9/8 Plux+RBS-lysis2-DT11011CP
9/8 Ptet+RBS-lacZα-DT11011CP
9/8 Ptet-pT181 anti+spinach-DT11011CP
9/8 Pcon-pT181 attenuater+DT11011CP

Electrophoresis

じっけんしゃ

LaneSampleEnzyme1Enzyme2
（°Д°）
LaneSampleEnzyme1Enzyme2
（°Д°）
LaneSampleEnzyme1Enzyme2
（°Д°）

Gel Extraction

じっけんしゃ

LaneDNAEnzyme
（＞ω・）b

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

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

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

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

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

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

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

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

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
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◎）　　　　

Gel Extraction

じっけんしゃ

LaneDNAEnzyme
（＞ω・）b

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

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

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

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