Template:Kyoto/Notebook/Aug 22
From 2013.igem.org
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(→Electrophoresis) |
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</div> | </div> | ||
+ | ===Restriction Enzyme Digestion=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Tatsui, Kojima, Honda and Ashida</span> | ||
+ | {| class="wikitable" | ||
+ | ! ||Fusion1 antisense (1) (300 µg/mL)||EcoRI||SpeI||buffer H||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3.3||1||1||3||21.7||30 | ||
+ | |- | ||
+ | |1 cut||0.7||0.2||0||1||8.1||10 | ||
+ | |- | ||
+ | |1 cut||0.7||0||0.2||1||8.1||10 | ||
+ | |- | ||
+ | |NC||0.7||0||0||1||8.3||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||Fusion1 antisense (1) (300 µg/mL)||XbaI||PstI||buffer M||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3.3||1||1||3||3||18.7||30 | ||
+ | |- | ||
+ | |1 cut||0.7||0.2||0||1||1||7.1||10 | ||
+ | |- | ||
+ | |1 cut||0.7||0||0.2||1||1||7.1||10 | ||
+ | |- | ||
+ | |NC||0.7||0||0||1||1||7.3||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||Fusion6 antisense (1) (336 µg/mL)||EcoRI||SpeI||buffer H||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3||1||1||3||22||30 | ||
+ | |- | ||
+ | |1 cut||0.6||0.2||0||1||8.2||10 | ||
+ | |- | ||
+ | |1 cut||0.6||0||0.2||1||8.2||10 | ||
+ | |- | ||
+ | |NC||0.6||0||0||1||8.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||Fusion6 antisense (1) (336 µg/mL)||XbaI||PstI||buffer M||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3||1||1||3||3||19||30 | ||
+ | |- | ||
+ | |1 cut||0.6||0.2||0||1||1||7.2||10 | ||
+ | |- | ||
+ | |1 cut||0.6||0||0.2||1||1||7.2||10 | ||
+ | |- | ||
+ | |NC||0.6||0||0||1||1||7.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||pT181 antisense (1) (178 µg/mL)||EcoRI||SpeI||buffer H||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||5.6||1||1||3||19.4||30 | ||
+ | |- | ||
+ | |1 cut||1.1||0.2||0||1||7.7||10 | ||
+ | |- | ||
+ | |1 cut||1.1||0||0.2||1||7.7||10 | ||
+ | |- | ||
+ | |NC||1.1||0||0||1||7.9||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||pT181 antisense (1) (178 µg/mL)||XbaI||PstI||buffer M||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||5.6||1||1||3||3||16.4||30 | ||
+ | |- | ||
+ | |1 cut||1.1||0.2||0||1||1||6.7||10 | ||
+ | |- | ||
+ | |1 cut||1.1||0||0.2||1||1||6.7||10 | ||
+ | |- | ||
+ | |NC||1.1||0||0||1||1||6.9||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||pT181 attenuator (1)||EcoRI||SpeI||buffer H||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3||1||1||3||22||30 | ||
+ | |- | ||
+ | |1 cut||0.6||0.2||0||1||8.2||10 | ||
+ | |- | ||
+ | |1 cut||0.6||0||0.2||1||8.2||10 | ||
+ | |- | ||
+ | |NC||0.6||0||0||1||8.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||pT181 attenuator (1)||XbaI||PstI||buffer M||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3||1||1||3||3||19||30 | ||
+ | |- | ||
+ | |1 cut||0.6||0.2||0||1||1||7.2||10 | ||
+ | |- | ||
+ | |1 cut||0.6||0||0.2||1||1||7.2||10 | ||
+ | |- | ||
+ | |NC||0.6||0||0||1||1||7.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||Fusion1 attenuator (1)||EcoRI||SpeI||buffer H||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3.3||1||1||3||21.7||30 | ||
+ | |- | ||
+ | |1 cut||0.7||0.2||0||1||8.1||10 | ||
+ | |- | ||
+ | |1 cut||0.7||0||0.2||1||8.1||10 | ||
+ | |- | ||
+ | |NC||0.7||0||0||1||8.3||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||Fusion1 attenuator (1)||XbaI||PstI||buffer M||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3.3||1||1||3||3||18.7||30 | ||
+ | |- | ||
+ | |1 cut||0.7||0.2||0||1||1||7.1||10 | ||
+ | |- | ||
+ | |1 cut||0.7||0||0.2||1||1||7.1||10 | ||
+ | |- | ||
+ | |NC||0.7||0||0||1||1||7.3||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||Fusion3m2 attenuator (1)||EcoRI||SpeI||buffer H||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3||1||1||3||22||30 | ||
+ | |- | ||
+ | |1 cut||0.6||0.2||0||1||8.2||10 | ||
+ | |- | ||
+ | |1 cut||0.6||0||0.2||1||8.2||10 | ||
+ | |- | ||
+ | |NC||0.6||0||0||1||8.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||Fusion3m2 attenuator (1)||XbaI||PstI||buffer M||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||3||1||1||3||3||19||30 | ||
+ | |- | ||
+ | |1 cut||0.6||0.2||0||1||1||7.2||10 | ||
+ | |- | ||
+ | |1 cut||0.6||0||0.2||1||1||7.2||10 | ||
+ | |- | ||
+ | |NC||0.6||0||0||1||1||7.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||Spinach (2)||EcoRI||SpeI||buffer B||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||2.3||0.5||0.5||3||0.3||23.4||30 | ||
+ | |- | ||
+ | |1 cut||0.5||0.1||0||1||0.1||8.3||10 | ||
+ | |- | ||
+ | |1 cut||0.5||0||0.1||1||0.1||8.3||10 | ||
+ | |- | ||
+ | |NC||0.5||0||0||1||0.1||8.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||tetR aptamer 12_P (2)||EcoRI||SpeI||buffer B||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||2.7||0.5||0.5||3||0.3||23||30 | ||
+ | |- | ||
+ | |1 cut||0.5||0.1||0||1||0.1||8.3||10 | ||
+ | |- | ||
+ | |1 cut||0.5||0||0.1||1||0.1||8.3||10 | ||
+ | |- | ||
+ | |NC||0.5||0||0||1||0.1||8.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||tetR aptamer 12_1R (2)||EcoRI||SpeI||buffer B||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||2.6||0.5||0.5||3||0.3||23.1||30 | ||
+ | |- | ||
+ | |1 cut||0.5||0.1||0||1||0.1||8.3||10 | ||
+ | |- | ||
+ | |1 cut||0.5||0||0.1||1||0.1||8.3||10 | ||
+ | |- | ||
+ | |NC||0.5||0||0||1||0.1||8.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||tetR aptamer 12_1M (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||2.7||0.5||0.5||3||0.3||23||30 | ||
+ | |- | ||
+ | |1 cut||0.5||0.1||0||1||0.1||8.3||10 | ||
+ | |- | ||
+ | |1 cut||0.5||0||0.1||1||0.1||8.3||10 | ||
+ | |- | ||
+ | |NC||0.5||0||0||1||0.1||8.4||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||J23100||SpeI||PstI||buffer B||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||7||0.5||0.5||3||0.3||18.7||30 | ||
+ | |- | ||
+ | |1 cut||1.4||0.1||0||1||0.1||7.4||10 | ||
+ | |- | ||
+ | |1 cut||1.4||0||0.1||1||0.1||7.4||10 | ||
+ | |- | ||
+ | |NC||1.4||0||0||1||0.1||7.5||10 | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | ! ||Plac (2)||SpeI||PstI||buffer B||BSA||MilliQ||total | ||
+ | |- | ||
+ | |2 cuts||11.2||0.5||0.5||3||0.3||14.5||30 | ||
+ | |- | ||
+ | |1 cut||2.2||0.1||0||1||0.1||6.6||10 | ||
+ | |- | ||
+ | |1 cut||2.2||0||0.1||1||0.1||6.6||10 | ||
+ | |- | ||
+ | |NC||2.2||0||0||1||0.1||6.7||10 | ||
+ | |} | ||
+ | </div> | ||
===Electrophoresis=== | ===Electrophoresis=== | ||
<div class="experiment"> | <div class="experiment"> | ||
- | <span class="author"> | + | <span class="author">No name</span> |
{| class="wikitable" | {| class="wikitable" | ||
!Lane||Sample | !Lane||Sample | ||
Line 130: | Line 332: | ||
|6||RBS(NC) -(1) | |6||RBS(NC) -(1) | ||
|- | |- | ||
- | |7||RBS(NC) -( | + | |7||RBS(NC) -(2) |
|- | |- | ||
|8||NC | |8||NC | ||
|} | |} | ||
- | [[File: | + | [[File:Igku Aug22 Electrophoresis(ColoP)-1.jpg]]<br> |
{| class="wikitable" | {| class="wikitable" | ||
!Lane||Sample | !Lane||Sample | ||
Line 156: | Line 358: | ||
|9||NC | |9||NC | ||
|} | |} | ||
- | [[File: | + | [[File:Igku Aug22 Electrophoresis(ColoP)-2.jpg]]<br> |
{| class="wikitable" | {| class="wikitable" | ||
!Lane||Sample | !Lane||Sample | ||
Line 174: | Line 376: | ||
|7||1kb ladder | |7||1kb ladder | ||
|} | |} | ||
- | [[File: | + | [[File:Igku Aug22 Electrophoresis(ColoP)-3.jpg]]<br> |
+ | </div> | ||
+ | |||
+ | ===Miniprep=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Ashida</span> | ||
+ | {|class="wikitable" | ||
+ | !DNA||concentration[µg/mL]||260/280||260/230 | ||
+ | |- | ||
+ | |8/22 RBS-lysis (1)||128||1.77||2.29 | ||
+ | |- | ||
+ | |8/22 RBS-lysis (2)||96||1.80||2.07 | ||
+ | |- | ||
+ | |8/22 pSB1C3 (1)||108||1.55||1.45 | ||
+ | |- | ||
+ | |8/22 pSB1C3 (2)||130||1.69||1.76 | ||
+ | |} | ||
</div> | </div> | ||
Latest revision as of 14:49, 25 September 2013
Contents |
Aug 22
Liquid culture
Sample | Medium |
---|---|
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (1) | 8/21 Plusgrow medium (+Amp) |
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (2) | 8/21 Plusgrow medium (+Amp) |
8/21 Pcon-RBS-GFP-DT control (1) | 8/21 Plusgrow medium (+Amp) |
8/21 Pcon-RBS-GFP-DT control (2) | 8/21 Plusgrow medium (+Amp) |
8/21 RBS-lysis1 (1) | 8/21 Plusgrow medium (+Amp) |
8/21 RBS-lysis1 (2) | 8/21 Plusgrow medium (+Amp) |
8/21 RBS control (1) | 8/21 Plusgrow medium (+Amp) |
8/21 RBS control (2) | 8/21 Plusgrow medium (+Amp) |
8/21 Ptet(pm)-RBS-tetR-DT (1) | 8/21 Plusgrow medium (+Amp) |
8/21 Ptet(pm)-RBS-tetR-DT (2) | 8/21 Plusgrow medium (+Amp) |
8/21 Ptet(pm) control (1) | 8/21 Plusgrow medium (+Amp) |
8/21 Ptet(pm) control (2) | 8/21 Plusgrow medium (+Amp) |
8/21 Plux-RBS-GFP-DT (1) | 8/21 Plusgrow medium (+Amp) |
8/21 pSB1C3(BBa_J04450) (1) | 8/21 Plusgrow medium (+Amp) |
8/21 pSB1C3(BBa_J04450) (2) | 8/21 Plusgrow medium (+Amp) |
incubate 37°C 10hour
Colony PCR
Sample | base pair |
---|---|
8/21 RBS-lysis1(1) | 400 |
8/21 RBS-lysis1(2) | 400 |
8/21 RBS control(1) | -- |
8/21 RBS control(2) | -- |
negative control | -- |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5min | 30s | 30s | 30s | 30cycles |
Sample | base pair |
---|---|
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (1) | 2143 |
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (2) | 2143 |
8/21 Pcon-RBS-luxR control(1) | -- |
8/21 Pcon-RBS-luxR control(2) | -- |
negative control | -- |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5min | 30s | 30s | 2min | 30cycles |
Sample | base pair |
---|---|
8/21 Ptet-RBS-tetR-DT (1) | 1216 |
8/21 Ptet-RBS-tetR-DT (2) | 1216 |
8/21 Ptet control (1) | -- |
8/21 Ptet control (2) | -- |
8/21 Plux-RBS-GFP-DT (1) | 1227 |
8/21 pSB1C3(BBa_J04450) (1) | 1353 |
8/21 pSB1C3(BBa_J04450) (2) | 1353 |
negative control | -- |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5min | 30s | 30s | 1min | 30cycles |
Restriction Enzyme Digestion
Fusion1 antisense (1) (300 µg/mL) | EcoRI | SpeI | buffer H | MilliQ | total | |
---|---|---|---|---|---|---|
2 cuts | 3.3 | 1 | 1 | 3 | 21.7 | 30 |
1 cut | 0.7 | 0.2 | 0 | 1 | 8.1 | 10 |
1 cut | 0.7 | 0 | 0.2 | 1 | 8.1 | 10 |
NC | 0.7 | 0 | 0 | 1 | 8.3 | 10 |
Fusion1 antisense (1) (300 µg/mL) | XbaI | PstI | buffer M | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 3.3 | 1 | 1 | 3 | 3 | 18.7 | 30 |
1 cut | 0.7 | 0.2 | 0 | 1 | 1 | 7.1 | 10 |
1 cut | 0.7 | 0 | 0.2 | 1 | 1 | 7.1 | 10 |
NC | 0.7 | 0 | 0 | 1 | 1 | 7.3 | 10 |
Fusion6 antisense (1) (336 µg/mL) | EcoRI | SpeI | buffer H | MilliQ | total | |
---|---|---|---|---|---|---|
2 cuts | 3 | 1 | 1 | 3 | 22 | 30 |
1 cut | 0.6 | 0.2 | 0 | 1 | 8.2 | 10 |
1 cut | 0.6 | 0 | 0.2 | 1 | 8.2 | 10 |
NC | 0.6 | 0 | 0 | 1 | 8.4 | 10 |
Fusion6 antisense (1) (336 µg/mL) | XbaI | PstI | buffer M | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 3 | 1 | 1 | 3 | 3 | 19 | 30 |
1 cut | 0.6 | 0.2 | 0 | 1 | 1 | 7.2 | 10 |
1 cut | 0.6 | 0 | 0.2 | 1 | 1 | 7.2 | 10 |
NC | 0.6 | 0 | 0 | 1 | 1 | 7.4 | 10 |
pT181 antisense (1) (178 µg/mL) | EcoRI | SpeI | buffer H | MilliQ | total | |
---|---|---|---|---|---|---|
2 cuts | 5.6 | 1 | 1 | 3 | 19.4 | 30 |
1 cut | 1.1 | 0.2 | 0 | 1 | 7.7 | 10 |
1 cut | 1.1 | 0 | 0.2 | 1 | 7.7 | 10 |
NC | 1.1 | 0 | 0 | 1 | 7.9 | 10 |
pT181 antisense (1) (178 µg/mL) | XbaI | PstI | buffer M | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 5.6 | 1 | 1 | 3 | 3 | 16.4 | 30 |
1 cut | 1.1 | 0.2 | 0 | 1 | 1 | 6.7 | 10 |
1 cut | 1.1 | 0 | 0.2 | 1 | 1 | 6.7 | 10 |
NC | 1.1 | 0 | 0 | 1 | 1 | 6.9 | 10 |
pT181 attenuator (1) | EcoRI | SpeI | buffer H | MilliQ | total | |
---|---|---|---|---|---|---|
2 cuts | 3 | 1 | 1 | 3 | 22 | 30 |
1 cut | 0.6 | 0.2 | 0 | 1 | 8.2 | 10 |
1 cut | 0.6 | 0 | 0.2 | 1 | 8.2 | 10 |
NC | 0.6 | 0 | 0 | 1 | 8.4 | 10 |
pT181 attenuator (1) | XbaI | PstI | buffer M | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 3 | 1 | 1 | 3 | 3 | 19 | 30 |
1 cut | 0.6 | 0.2 | 0 | 1 | 1 | 7.2 | 10 |
1 cut | 0.6 | 0 | 0.2 | 1 | 1 | 7.2 | 10 |
NC | 0.6 | 0 | 0 | 1 | 1 | 7.4 | 10 |
Fusion1 attenuator (1) | EcoRI | SpeI | buffer H | MilliQ | total | |
---|---|---|---|---|---|---|
2 cuts | 3.3 | 1 | 1 | 3 | 21.7 | 30 |
1 cut | 0.7 | 0.2 | 0 | 1 | 8.1 | 10 |
1 cut | 0.7 | 0 | 0.2 | 1 | 8.1 | 10 |
NC | 0.7 | 0 | 0 | 1 | 8.3 | 10 |
Fusion1 attenuator (1) | XbaI | PstI | buffer M | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 3.3 | 1 | 1 | 3 | 3 | 18.7 | 30 |
1 cut | 0.7 | 0.2 | 0 | 1 | 1 | 7.1 | 10 |
1 cut | 0.7 | 0 | 0.2 | 1 | 1 | 7.1 | 10 |
NC | 0.7 | 0 | 0 | 1 | 1 | 7.3 | 10 |
Fusion3m2 attenuator (1) | EcoRI | SpeI | buffer H | MilliQ | total | |
---|---|---|---|---|---|---|
2 cuts | 3 | 1 | 1 | 3 | 22 | 30 |
1 cut | 0.6 | 0.2 | 0 | 1 | 8.2 | 10 |
1 cut | 0.6 | 0 | 0.2 | 1 | 8.2 | 10 |
NC | 0.6 | 0 | 0 | 1 | 8.4 | 10 |
Fusion3m2 attenuator (1) | XbaI | PstI | buffer M | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 3 | 1 | 1 | 3 | 3 | 19 | 30 |
1 cut | 0.6 | 0.2 | 0 | 1 | 1 | 7.2 | 10 |
1 cut | 0.6 | 0 | 0.2 | 1 | 1 | 7.2 | 10 |
NC | 0.6 | 0 | 0 | 1 | 1 | 7.4 | 10 |
Spinach (2) | EcoRI | SpeI | buffer B | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.3 | 0.5 | 0.5 | 3 | 0.3 | 23.4 | 30 |
1 cut | 0.5 | 0.1 | 0 | 1 | 0.1 | 8.3 | 10 |
1 cut | 0.5 | 0 | 0.1 | 1 | 0.1 | 8.3 | 10 |
NC | 0.5 | 0 | 0 | 1 | 0.1 | 8.4 | 10 |
tetR aptamer 12_P (2) | EcoRI | SpeI | buffer B | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.7 | 0.5 | 0.5 | 3 | 0.3 | 23 | 30 |
1 cut | 0.5 | 0.1 | 0 | 1 | 0.1 | 8.3 | 10 |
1 cut | 0.5 | 0 | 0.1 | 1 | 0.1 | 8.3 | 10 |
NC | 0.5 | 0 | 0 | 1 | 0.1 | 8.4 | 10 |
tetR aptamer 12_1R (2) | EcoRI | SpeI | buffer B | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.6 | 0.5 | 0.5 | 3 | 0.3 | 23.1 | 30 |
1 cut | 0.5 | 0.1 | 0 | 1 | 0.1 | 8.3 | 10 |
1 cut | 0.5 | 0 | 0.1 | 1 | 0.1 | 8.3 | 10 |
NC | 0.5 | 0 | 0 | 1 | 0.1 | 8.4 | 10 |
tetR aptamer 12_1M (1) | EcoRI | SpeI | buffer B | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.7 | 0.5 | 0.5 | 3 | 0.3 | 23 | 30 |
1 cut | 0.5 | 0.1 | 0 | 1 | 0.1 | 8.3 | 10 |
1 cut | 0.5 | 0 | 0.1 | 1 | 0.1 | 8.3 | 10 |
NC | 0.5 | 0 | 0 | 1 | 0.1 | 8.4 | 10 |
J23100 | SpeI | PstI | buffer B | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 7 | 0.5 | 0.5 | 3 | 0.3 | 18.7 | 30 |
1 cut | 1.4 | 0.1 | 0 | 1 | 0.1 | 7.4 | 10 |
1 cut | 1.4 | 0 | 0.1 | 1 | 0.1 | 7.4 | 10 |
NC | 1.4 | 0 | 0 | 1 | 0.1 | 7.5 | 10 |
Plac (2) | SpeI | PstI | buffer B | BSA | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 11.2 | 0.5 | 0.5 | 3 | 0.3 | 14.5 | 30 |
1 cut | 2.2 | 0.1 | 0 | 1 | 0.1 | 6.6 | 10 |
1 cut | 2.2 | 0 | 0.1 | 1 | 0.1 | 6.6 | 10 |
NC | 2.2 | 0 | 0 | 1 | 0.1 | 6.7 | 10 |
Electrophoresis
Lane | Sample |
---|---|
1 | 100bp ladder |
2 | RBS-lysis1 -(1) |
3 | RBS-lysis1 -(2) |
4 | RBS-lysis2 -(1) |
5 | RBS-lysis2 -(2) |
6 | RBS(NC) -(1) |
7 | RBS(NC) -(2) |
8 | NC |
Lane | Sample |
---|---|
1 | 1kb ladder |
2 | Ptet-RBS-tetR-DT -(1) |
3 | Ptet-RBS-tetR-DT -(2) |
4 | Ptet(NC) -(1) |
5 | Ptet(NC) -(2) |
6 | Plux-RBS-GFP-DT -(1) |
7 | pSB1C3 -(1) |
8 | pSB1C3 -(2) |
9 | NC |
Lane | Sample |
---|---|
1 | 1kb ladder |
2 | Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT -(1) |
3 | Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT -(2) |
4 | Pcon-RBS-luxR -(1) |
5 | Pcon-RBS-luxR -(2) |
6 | NC |
7 | 1kb ladder |
Miniprep
DNA | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
8/22 RBS-lysis (1) | 128 | 1.77 | 2.29 |
8/22 RBS-lysis (2) | 96 | 1.80 | 2.07 |
8/22 pSB1C3 (1) | 108 | 1.55 | 1.45 |
8/22 pSB1C3 (2) | 130 | 1.69 | 1.76 |
Electrophoresis
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1 | 100bp | -- | -- |
2 | 8/22 pT181 attenuator (1) | EcoRI | SpeI |
3 | 8/22 pT181 attenuator (1) | EcoRI | -- |
4 | 8/22 pT181 attenuator (1) | -- | SpeI |
5 | 8/22 pT181 attenuator (1) | -- | -- |
6 | 8/22 pT181 attenuator (1) | XbaI | PstI |
7 | 8/22 pT181 attenuator (1) | XbaI | -- |
8 | 8/22 pT181 attenuator (1) | -- | PstI |
9 | 8/22 pT181 attenuator (1) | -- | -- |
10 | 8/22 Fusion6 antisense (1) | EcoRI | SpeI |
11 | 8/22 Fusion6 antisense (1) | EcoRI | -- |
12 | 8/22 Fusion6 antisense (1) | -- | SpeI |
13 | 8/22 Fusion6 antisense (1) | -- | -- |
14 | 100bp | -- | -- |
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1 | 100bp | -- | -- |
2 | 8/22 Fusion6 antisense (1) | XbaI | PstI |
3 | 8/22 Fusion6 antisense (1) | XbaI | -- |
4 | 8/22 Fusion6 antisense (1) | -- | PstI |
5 | 8/22 Fusion6 antisense (1) | -- | -- |
6 | 8/22 Fusion1 antisense (1) | EcoRI | SpeI |
7 | 8/22 Fusion1 antisense (1) | EcoRI | -- |
8 | 8/22 Fusion1 antisense (1) | -- | SpeI |
9 | 8/22 Fusion1 antisense (1) | -- | -- |
10 | 8/22 Fusion1 antisense (1) | XbaI | PstI |
11 | 8/22 Fusion1 antisense (1) | XbaI | -- |
12 | 8/22 Fusion1 antisense (1) | -- | PstI |
13 | 8/22 Fusion1 antisense (1) | -- | -- |
14 | 100bp | -- | -- |