Template:Kyoto/Notebook/Sep 8

From 2013.igem.org

(Difference between revisions)
(→Restriction Enzyme Digestion)
(→Restriction Enzyme Digestion)
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! ||8/21 F3m2 atte||E||S||X||P||BSA||Buffer||MilliQ||total
! ||8/21 F3m2 atte||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|2 cuts||6.0µL||0µL||0µL||1µL||1µL||3µL||3µL||16µL||30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|NC|| 0.3µL||0µL||0µL|| 0µL|| 0µL||1µL||1µL||7.7µL||10µL
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||8/21 F1 anti||E||S||X||P||BSA||Buffer||MilliQ||total
! ||8/21 F1 anti||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|2 cuts||6.7µL||0µL||0µL||1µL||1µL||3µL||3µL||15.3µL||30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|NC|| 0.3µL||0µL||0µL|| 0µL|| 0µL||1µL||1µL||7.7µL||10µL
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||8/21 F6 anti||E||S||X||P||BSA||Buffer||MilliQ||total
! ||8/21 F6 anti||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|2 cuts||6.0µL||0µL||0µL||1µL||1µL||3µL||3µL||16µL||30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|NC|| 0.3µL||0µL||0µL|| 0µL|| 0µL||1µL||1µL||7.7µL||10µL
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||8/21 tetR aptamer 12_1M||E||S||X||P||BSA||Buffer||MilliQ||total
! ||8/21 tetR aptamer 12_1M||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|2 cuts||5.3µL||1µL||1µL||0µL||0µL||3µL||3µL||16.7µL||30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|NC|| 0.3µL||0µL||0µL|| 0µL|| 0µL||1µL||1µL||7.7µL||10µL
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||8/21 tetR aptamer 12_P||E||S||X||P||BSA||Buffer||MilliQ||total
! ||8/21 tetR aptamer 12_P||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|2 cuts||13.3µL||1µL||1µL||0µL||0µL||3µL||3µL||8.7µL||30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|NC||0.7µL||0µL||0µL||0µL||0µL||1µL||1µL||7.3µL||10µL
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||9/4 spin-DT||E||S||X||P||BSA||Buffer||MilliQ||total
! ||9/4 spin-DT||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
||2 cuts||5.4µL||0µL||0µL||1µL||1µL||3µL||3µL||16.6µL||30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|NC||0.3µL||0µL||0µL||0µL||0µL||1µL||1µL||7.7µL||10µL
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||9/7 RBS-lysis1-DT||E||S||X||P||BSA||Buffer||MilliQ||total
! ||9/7 RBS-lysis1-DT||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|2 cuts|||10µL||0µL||0µL||1µL||1µL||3µL||3µL||12µL||30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|NC||0.5µL||0µL||0µL||0µL||0µL||1µL||1µL||7.5µL||10µL
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||9/6 RBS-lysis2-DT||E||S||X||P||BSA||Buffer||MilliQ||total
! ||9/6 RBS-lysis2-DT||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
||2 cuts||13µL||0µL||0µL||1µL||1µL||3µL||3µL||9µL||30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|NC||0.7µL||0µL||0µL||0µL||0µL||1µL||1µL||7.3µL||10µL
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||9/7 RBS-lysis3-DT||E||S||X||P||BSA||Buffer||MilliQ||total
! ||9/7 RBS-lysis3-DT||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|2 cuts|||10µL||0µL||0µL||1µL||1µL||3µL||3µL||12µL||30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|NC||0.5µL||0µL||0µL||0µL||0µL||1µL||1µL||7.5µL||10µL
 +
 
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||8/20 J23100 RBS-lacZα-DT-(1)||E||S||X||P||BSA||Buffer||MilliQ||total
! ||8/20 J23100 RBS-lacZα-DT-(1)||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|2 cuts|||7µL||0.5µL||0.5µL||0µL||0µL||3µL||3µL||16µL||30µL
 +
|-
 +
|NC||0.5µL||0µL||0µL||0µL||0µL||1µL||1µL||7.5µL||10µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
 
|}
|}
{| class="wikitable"
{| class="wikitable"
! ||8/20 RBS-lacZα-DT||E||S||X||P||BSA||Buffer||MilliQ||total
! ||8/20 RBS-lacZα-DT||E||S||X||P||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
+
|2 cuts||10µL||0.5µL||0.5µL||0µL||0µL||3µL||3µL||13µL||30µL
 +
|-
 +
|NC||0.5µL||0µL||0µL||0µL||0µL||1µL||1µL||7.5µL||10µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL
 
|}
|}

Revision as of 01:00, 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

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

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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