Template:Kyoto/Notebook/Aug 30

From 2013.igem.org

(Difference between revisions)
(Created page with "==Aug 30== ===Gel Extraction===")
(→Miniprep)
 
(34 intermediate revisions not shown)
Line 1: Line 1:
==Aug 30==
==Aug 30==
===Gel Extraction===
===Gel Extraction===
 +
<div class="experiment">
 +
<span class="author">No Name</span>
 +
{| class="wikitable"
 +
!Name||quantity||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|pT181 attenuator(2) (EcoRI & SpeI)||305.3 ||2.2 ||1.66 ||0.36
 +
|-
 +
|pT181 attenuator(2) (XbaI & PstI)||217.7 ||3.9 ||1.73 ||0.04
 +
|-
 +
|pT181 antisense(2) (EcoRI & SpeI)||347.7 ||3.1 ||1.88 ||0.31
 +
|-
 +
|pT181 antisense(2) (XbaI & PstI)||376.5 ||5.9 ||0.49 ||2.36
 +
|-
 +
|pSB1C3 (EcoRI & SpeI)||436.6 ||30.7 ||1.03 ||1.28
 +
|-
 +
|pSB1C3 (XbaI & PstI)||457.4 ||7.9 ||1.65 ||0.42
 +
|-
 +
|Spinach (EcoRI & SpeI)||490.5 ||3.0 ||1.87 ||0.26
 +
|-
 +
|tetR aptamer 12_1R (EcoRI & SpeI)||451.7 ||-12.8 ||5.56 ||-0.04
 +
|}
 +
</div>
 +
 +
===Miniprep===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{|class="wikitable"
 +
!DNA||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|RBS-lysis2 9||264.6||1.97||2.21
 +
|-
 +
|RBS-lysis2 12||220.0||1.89||2.46
 +
|-
 +
|RBS-lysis2 14||195.3||1.88||1.53
 +
|}
 +
</div>
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT(8/20 LB Amp) (1)~(4)||2143
 +
|-
 +
|8/29 Plux-RBS-GFP-DT(8/27 LB-CP) (1)~(2)||1227
 +
|-
 +
|8/29 Pbad/araC-RBS-RFP{BBa_I13516} (8/21 LB-CP) (1)~(2)||2256
 +
|-
 +
|8/29 RBS-lysis3-DT-(1)||1210
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||2min||30cycle
 +
|}
 +
[[File:Igku_Aug30_colonyPCR_N1.jpg]]
 +
</div>
 +
 +
===Ethanol Precipitation===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{|class="wikitable"
 +
!DNA
 +
|-
 +
|pT181 attenuator-(2) (EcoRI & SpeI)
 +
|-
 +
|pT181 attenuator-(2) (XbaI & PstI)
 +
|-
 +
|pT181 antisense-(2) (EcoRI & SpeI)
 +
|-
 +
|pT181 antisense-(2) (XbaI & PstI)
 +
|-
 +
|pSB1C3 (EcoRI & SpeI)
 +
|-
 +
|pSB1C3 (XbaI & PstI)
 +
|-
 +
|Spinach (EcoRI & SpeI)
 +
|-
 +
|tetR aptamer 12_1R (EcoRI & SpeI)
 +
|}
 +
</div>
 +
 +
===Restriction Enzyme Digestion===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
! ||8/30 RBS-lysis2 (9)||EcoRI||SpeI||10x bufferB||100x BSA||MilliQ||total
 +
|-
 +
|2 cuts||7.5 &micro;L||0.5 &micro;L||0.5 &micro;L||3 &micro;L||0.3 &micro;L||18.2 &micro;L||30 &micro;L
 +
|-
 +
|NC||0.5 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||0.1 &micro;L||8.4 &micro;L||10 &micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/22 pSB1C3-(2)||EcoRI||SpeI||10x Buffer||100x BSA||MilliQ||total
 +
|-
 +
|2 cuts||8 &micro;L||0.5 &micro;L||0.5 &micro;L||3 &micro;L||0.3 &micro;L||17.7 &micro;L||30 &micro;L
 +
|-
 +
|NC||1 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||0.1 &micro;L||7.9 &micro;L||10 &micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/22 pSB1C3-(2)||XbaI||PstI||BufferD||BSA||MilliQ||total
 +
|-
 +
|2 cuts||8 &micro;L||0.5 &micro;L||0.5 &micro;L||3 &micro;L||0.3 &micro;L||17.7 &micro;L||30 &micro;L
 +
|-
 +
|NC||1 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||0.1 &micro;L||7.9 &micro;L||10 &micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 Spinach (1)||EcoRI||SpeI||10x BufferB||100x BSA||MilliQ||total
 +
|-
 +
|2 cuts||4 &micro;L||0.5 &micro;L||0.5 &micro;L||3 &micro;L||0.3 &micro;L||21.9 &micro;L||30 &micro;L
 +
|-
 +
|NC||0.5 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||0.1 &micro;L||8.4 &micro;L||10 &micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 tetR aptamer 12_1R (1)||EcoRI||SpeI||10xBufferB||100xBSA||MilliQ||total
 +
|-
 +
|2 cuts||3 &micro;L||0.5 &micro;L||0.5 &micro;L||3 &micro;L||0.3 &micro;L||22.7 &micro;L||30 &micro;L
 +
|-
 +
|NC||0.4 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||0.1 &micro;L||8.4 &micro;L||10 &micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 pT181 attenuator-(2)||EcoRI||SpeI||XbaI||PstI||10x BufferB||10x BufferD||100x BSA||MilliQ||total
 +
|-
 +
|2 cuts||4 &micro;L||0.5 &micro;L||0.5 &micro;L||0 &micro;L||0 &micro;L||3 &micro;L||0 &micro;L||0.3 &micro;L||21.7 &micro;L||30 &micro;L
 +
|-
 +
|NC||0.5 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||0 &micro;L||0.1 &micro;L||8.4 &micro;L||10 &micro;L
 +
|-
 +
|2 cuts||4 &micro;L||0 &micro;L||0 &micro;L||0.5 &micro;L||0.5 &micro;L||0 &micro;L||3 &micro;L||0.3 &micro;L||21.7 &micro;L||30 &micro;L
 +
|-
 +
|NC||0.5 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||0.1 &micro;L||8.4 &micro;L||10 &micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 pT181 antisense-(2)||EcoRI||SpeI||XbaI||PstI||10x BufferB||10x BufferD||100x BSA||MilliQ||total
 +
|-
 +
|2 cuts||9 &micro;L||0.5 &micro;L||0.5 &micro;L||0 &micro;L||0 &micro;L||3 &micro;L||0 &micro;L||0.3 &micro;L||16.7 &micro;L||30 &micro;L
 +
|-
 +
|NC||1 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||0 &micro;L||0.1 &micro;L||7.9 &micro;L||10 &micro;L
 +
|-
 +
|2 cuts||9 &micro;L||0 &micro;L||0 &micro;L||0.5 &micro;L||0.5 &micro;L||0 &micro;L||3 &micro;L||0.3 &micro;L||16.7 &micro;L||30 &micro;L
 +
|-
 +
|NC||1 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||0.1 &micro;L||7.9 &micro;L||10 &micro;L
 +
|}
 +
</div>
 +
===Electrophoresis===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp ladder||--||--
 +
|-
 +
|2||RBS-lysis2-(9)||EcoRI||SpeI
 +
|-
 +
|3||RBS-lysis2-(9)||--||--
 +
|-
 +
|4||pSB1C3-(2)||EcoRI||SpeI
 +
|-
 +
|5||pSB1C3-(2)||--||--
 +
|-
 +
|6||pSB1C3-(2)||XbaI||PstI
 +
|-
 +
|7||pSB1C3-(2)||--||--
 +
|-
 +
|8||tRNA-Spinach-(1)||EcoRI||SpeI
 +
|-
 +
|9||tRNA-Spinach-(1)||--||--
 +
|-
 +
|10||tetR aptamer12_1R||EcoRI||SpeI
 +
|-
 +
|11||tetR aptamer12_1R||--||--
 +
|-
 +
|12||pT181 attenuator||EcoRI||SpeI
 +
|-
 +
|13||pT181 attenuator||EcoRI||SpeI
 +
|-
 +
|14||100bp ladder||--||--
 +
|}
 +
[[File:Igku Aug30 Electrophoresis(N2-1).jpg]]<br>
 +
 +
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp ladder||--||--
 +
|-
 +
|2||pT181 antisense||EcoRI||SpeI
 +
|-
 +
|3||pT181 antisense||--||--
 +
|-
 +
|4||pT181 antisense||XbaI||PstI
 +
|-
 +
|5||pT181 antisense||--||--
 +
|-
 +
|6||pT181 attenuator||XbaI||PstI
 +
|-
 +
|7||pT181 attenuator||--||--
 +
|-
 +
|8||100bp ladder||--||--
 +
|}
 +
[[File:Igku Aug30 Electrophoresis(N2-2).jpg]]<br>
 +
 +
{| class="wikitable"
 +
!Lane||Sample
 +
|-
 +
|1||1kb ladder
 +
|-
 +
|2||8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)(Colony PCR product)
 +
|-
 +
|3||8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)(Colony PCR product)
 +
|-
 +
|4||8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)(Colony PCR product)
 +
|-
 +
|5||8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)(Colony PCR product)
 +
|-
 +
|6||Pbad/araC-RBS-RFP-DT-(1)(Colony PCR product)
 +
|-
 +
|7||Pbad-araC-RBS-RFP-DT-(2)(Colony PCR product)
 +
|}
 +
[[File:Igku Aug30 Electrophoresis(N2-3).jpg]]<br>
 +
 +
===Liquid Culture===
 +
<!-- ここから -->
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)||Plusgrow medium(+Amp)
 +
|-
 +
|8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)||Plusgrow medium(+Amp)
 +
|-
 +
|8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)||Plusgrow medium(+Amp)
 +
|-
 +
|8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)||Plusgrow medium(+Amp)
 +
|-
 +
|8/29 Plux-RBS-GFP-DT-(1)||Plusgrow medium(+CP)
 +
|-
 +
|8/29 Plux-RBS-GFP-DT-(2)||Plusgrow medium(+CP)
 +
|-
 +
|8/29 Pbad/araC-RBS-RFP-DT-(1)||Plusgrow medium(+CP)
 +
|-
 +
|8/29 Pbad-araC-RBS-RFP-DT-(2)||Plusgrow medium(+CP)
 +
|-
 +
|RBS-lysis3-DT-(1)||Plusgrow medium(+CP)
 +
|}
 +
</div>

Latest revision as of 03:49, 28 September 2013

Contents

Aug 30

Gel Extraction

No Name

Namequantityconcentration[µg/mL]260/280260/230
pT181 attenuator(2) (EcoRI & SpeI)305.3 2.2 1.66 0.36
pT181 attenuator(2) (XbaI & PstI)217.7 3.9 1.73 0.04
pT181 antisense(2) (EcoRI & SpeI)347.7 3.1 1.88 0.31
pT181 antisense(2) (XbaI & PstI)376.5 5.9 0.49 2.36
pSB1C3 (EcoRI & SpeI)436.6 30.7 1.03 1.28
pSB1C3 (XbaI & PstI)457.4 7.9 1.65 0.42
Spinach (EcoRI & SpeI)490.5 3.0 1.87 0.26
tetR aptamer 12_1R (EcoRI & SpeI)451.7 -12.8 5.56 -0.04

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
RBS-lysis2 9264.61.972.21
RBS-lysis2 12220.01.892.46
RBS-lysis2 14195.31.881.53

Colony PCR

No name

Samplebase pair
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT(8/20 LB Amp) (1)~(4)2143
8/29 Plux-RBS-GFP-DT(8/27 LB-CP) (1)~(2)1227
8/29 Pbad/araC-RBS-RFP{BBa_I13516} (8/21 LB-CP) (1)~(2)2256
8/29 RBS-lysis3-DT-(1)1210
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s2min30cycle

Igku Aug30 colonyPCR N1.jpg

Ethanol Precipitation

No name

DNA
pT181 attenuator-(2) (EcoRI & SpeI)
pT181 attenuator-(2) (XbaI & PstI)
pT181 antisense-(2) (EcoRI & SpeI)
pT181 antisense-(2) (XbaI & PstI)
pSB1C3 (EcoRI & SpeI)
pSB1C3 (XbaI & PstI)
Spinach (EcoRI & SpeI)
tetR aptamer 12_1R (EcoRI & SpeI)

Restriction Enzyme Digestion

No name

8/30 RBS-lysis2 (9)EcoRISpeI10x bufferB100x BSAMilliQtotal
2 cuts7.5 µL0.5 µL0.5 µL3 µL0.3 µL18.2 µL30 µL
NC0.5 µL0 µL0 µL1 µL0.1 µL8.4 µL10 µL
8/22 pSB1C3-(2)EcoRISpeI10x Buffer100x BSAMilliQtotal
2 cuts8 µL0.5 µL0.5 µL3 µL0.3 µL17.7 µL30 µL
NC1 µL0 µL0 µL1 µL0.1 µL7.9 µL10 µL
8/22 pSB1C3-(2)XbaIPstIBufferDBSAMilliQtotal
2 cuts8 µL0.5 µL0.5 µL3 µL0.3 µL17.7 µL30 µL
NC1 µL0 µL0 µL1 µL0.1 µL7.9 µL10 µL
8/21 Spinach (1)EcoRISpeI10x BufferB100x BSAMilliQtotal
2 cuts4 µL0.5 µL0.5 µL3 µL0.3 µL21.9 µL30 µL
NC0.5 µL0 µL0 µL1 µL0.1 µL8.4 µL10 µL
8/21 tetR aptamer 12_1R (1)EcoRISpeI10xBufferB100xBSAMilliQtotal
2 cuts3 µL0.5 µL0.5 µL3 µL0.3 µL22.7 µL30 µL
NC0.4 µL0 µL0 µL1 µL0.1 µL8.4 µL10 µL
8/21 pT181 attenuator-(2)EcoRISpeIXbaIPstI10x BufferB10x BufferD100x BSAMilliQtotal
2 cuts4 µL0.5 µL0.5 µL0 µL0 µL3 µL0 µL0.3 µL21.7 µL30 µL
NC0.5 µL0 µL0 µL0 µL0 µL1 µL0 µL0.1 µL8.4 µL10 µL
2 cuts4 µL0 µL0 µL0.5 µL0.5 µL0 µL3 µL0.3 µL21.7 µL30 µL
NC0.5 µL0 µL0 µL0 µL0 µL0 µL1 µL0.1 µL8.4 µL10 µL
8/21 pT181 antisense-(2)EcoRISpeIXbaIPstI10x BufferB10x BufferD100x BSAMilliQtotal
2 cuts9 µL0.5 µL0.5 µL0 µL0 µL3 µL0 µL0.3 µL16.7 µL30 µL
NC1 µL0 µL0 µL0 µL0 µL1 µL0 µL0.1 µL7.9 µL10 µL
2 cuts9 µL0 µL0 µL0.5 µL0.5 µL0 µL3 µL0.3 µL16.7 µL30 µL
NC1 µL0 µL0 µL0 µL0 µL0 µL1 µL0.1 µL7.9 µL10 µL

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp ladder----
2RBS-lysis2-(9)EcoRISpeI
3RBS-lysis2-(9)----
4pSB1C3-(2)EcoRISpeI
5pSB1C3-(2)----
6pSB1C3-(2)XbaIPstI
7pSB1C3-(2)----
8tRNA-Spinach-(1)EcoRISpeI
9tRNA-Spinach-(1)----
10tetR aptamer12_1REcoRISpeI
11tetR aptamer12_1R----
12pT181 attenuatorEcoRISpeI
13pT181 attenuatorEcoRISpeI
14100bp ladder----

Igku Aug30 Electrophoresis(N2-1).jpg


LaneSampleEnzyme1Enzyme2
1100bp ladder----
2pT181 antisenseEcoRISpeI
3pT181 antisense----
4pT181 antisenseXbaIPstI
5pT181 antisense----
6pT181 attenuatorXbaIPstI
7pT181 attenuator----
8100bp ladder----

Igku Aug30 Electrophoresis(N2-2).jpg

LaneSample
11kb ladder
28/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)(Colony PCR product)
38/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)(Colony PCR product)
48/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)(Colony PCR product)
58/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)(Colony PCR product)
6Pbad/araC-RBS-RFP-DT-(1)(Colony PCR product)
7Pbad-araC-RBS-RFP-DT-(2)(Colony PCR product)

Igku Aug30 Electrophoresis(N2-3).jpg

Liquid Culture

No name

Samplemedium
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)Plusgrow medium(+Amp)
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)Plusgrow medium(+Amp)
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)Plusgrow medium(+Amp)
8/29 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)Plusgrow medium(+Amp)
8/29 Plux-RBS-GFP-DT-(1)Plusgrow medium(+CP)
8/29 Plux-RBS-GFP-DT-(2)Plusgrow medium(+CP)
8/29 Pbad/araC-RBS-RFP-DT-(1)Plusgrow medium(+CP)
8/29 Pbad-araC-RBS-RFP-DT-(2)Plusgrow medium(+CP)
RBS-lysis3-DT-(1)Plusgrow medium(+CP)
Retrieved from "http://2013.igem.org/Template:Kyoto/Notebook/Aug_30"