Template:Kyoto/Notebook/Aug 17
From 2013.igem.org
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|8/15 BBa_K112806(lysis2) -(1)||828 | |8/15 BBa_K112806(lysis2) -(1)||828 | ||
|- | |- | ||
- | | | + | |NC||-- |
|} | |} | ||
{| class="wikitable" | {| class="wikitable" | ||
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|8/10 BBa_Q0400 -(1)||3995 | |8/10 BBa_Q0400 -(1)||3995 | ||
|- | |- | ||
- | | | + | |NC||-- |
|} | |} | ||
{| class="wikitable" | {| class="wikitable" | ||
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</div> | </div> | ||
+ | |||
===Electrophoresis=== | ===Electrophoresis=== | ||
<div class="experiment"> | <div class="experiment"> | ||
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|NC||1.3µL||--||--||1.0µL||7.7µL||10µL | |NC||1.3µL||--||--||1.0µL||7.7µL||10µL | ||
|} | |} | ||
- | |||
{| class="wikitable" | {| class="wikitable" | ||
! ||8/15 B0034(RBS) -(2)||SpeI||PstI||Buffer||MilliQ||total | ! ||8/15 B0034(RBS) -(2)||SpeI||PstI||Buffer||MilliQ||total | ||
Line 136: | Line 136: | ||
|NC||2.0µL||--||--||1.0µL||7.0µL||10µL | |NC||2.0µL||--||--||1.0µL||7.0µL||10µL | ||
|} | |} | ||
- | |||
{| class="wikitable" | {| class="wikitable" | ||
! ||8/17 K117000(lysis1) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ! ||8/17 K117000(lysis1) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ||
Line 148: | Line 147: | ||
|NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | |NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | ||
|} | |} | ||
- | |||
{| class="wikitable" | {| class="wikitable" | ||
! ||8/17 K748002(lysis3) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ! ||8/17 K748002(lysis3) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ||
Line 160: | Line 158: | ||
|NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | |NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | ||
|} | |} | ||
- | |||
{| class="wikitable" | {| class="wikitable" | ||
! ||8/17 I13504(RBS-GFP-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ! ||8/17 I13504(RBS-GFP-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ||
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|NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | |NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | ||
|} | |} | ||
- | |||
{| class="wikitable" | {| class="wikitable" | ||
! ||8/17 I0462(RBS-luxR-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ! ||8/17 I0462(RBS-luxR-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ||
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|NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | |NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | ||
|} | |} | ||
- | |||
{| class="wikitable" | {| class="wikitable" | ||
! ||8/17 I732020(RBS-lacZα-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ! ||8/17 I732020(RBS-lacZα-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total | ||
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|NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | |NC||1.0µL||--||--||1.0µL||1.0µL||7.0µL||10µL | ||
|} | |} | ||
- | |||
{| class="wikitable" | {| class="wikitable" | ||
! ||8/11 R6061 -(1)||SpeI||PstI||Buffer||MilliQ||total | ! ||8/11 R6061 -(1)||SpeI||PstI||Buffer||MilliQ||total | ||
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|NC||3.0µL||--||--||1.0µL||6.0µL||10µL | |NC||3.0µL||--||--||1.0µL||6.0µL||10µL | ||
|} | |} | ||
- | *at 37°C, for | + | *incubate at 37 °C, for 1 h |
</div> | </div> | ||
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|17||K748002(lysis3) -(1)||--||-- | |17||K748002(lysis3) -(1)||--||-- | ||
|} | |} | ||
- | [[File: | + | [[File:Igku Aug17 Electrophoresis(1).jpg]]<br> |
{| class="wikitable" | {| class="wikitable" | ||
!Lane||Sample||Enzyme1||Enzyme2 | !Lane||Sample||Enzyme1||Enzyme2 | ||
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|34||R0061(Plux) -(1)||--||-- | |34||R0061(Plux) -(1)||--||-- | ||
|} | |} | ||
- | [[File: | + | [[File:Igku Aug17 Electrophoresis(2).jpg]]<br> |
</div> | </div> | ||
Latest revision as of 07:41, 27 September 2013
Contents |
Aug 17
Colony PCR
Sample | base pair |
---|---|
8/15 BBa_K112806(lysis2) -(1) | 828 |
NC | -- |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5min | 30s | 30s | 1min | 30cycles |
Sample | base pair |
---|---|
8/10 BBa_Q0400 -(1) | 3995 |
NC | -- |
PreDenature | Denature | Annealing | Extension | cycle |
---|---|---|---|---|
94°C | 94°C | 55°C | 68°C | -- |
5min | 30s | 30s | 30s | 30cycles |
Electrophoresis
Liquid Culture
Sample | medium |
---|---|
8/15 BBa_K112806(lysis2) -(1) | Plusgrow medium(+CP) |
8/15 BBa_Q04400(tetR) -(1) | Plusgrow medium(+Amp) |
8/16 pBR322 -(1) | Plusgrow medium(+Amp) |
Master Plate
Number | Use LB plate (+CP) |
---|---|
1 | BBa_K112806(lysis2) -(1) |
Number | Use LB plate (+Kana) |
---|---|
1 | BBa_Q04400(tetR) -(1) |
Miniprep
DNA | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
K748002(lysis3) -(1) | 330 | 1.66 | 2.08 |
B0015(DT) -(1) | 188 | 1.73 | 1.82 |
R0061(Plux) -(1) | 76 | 1.64 | 1.30 |
K117000(lysis1) -(1) | 286 | 1.71 | 2.01 |
K117000(lysis1) -(2) | 248 | 1.69 | 1.78 |
I732020(RBS-lacZα-DT) -(1) | 198 | 1.71 | 1.93 |
I13504(RBS-GFP-DT) -(1) | 248 | 1.69 | 2.03 |
I13504(RBS-GFP-DT) -(2) | 246 | 1.55 | 1.73 |
I0462(RBS-luxR-DT) -(1) | 326 | 1.65 | 1.74 |
Restriction Enzyme Digestion
8/11 J23100 -(2) | SpeI | PstI | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|
2 cuts | 4.0µL | 1.0µL | 1.0µL | 3.0µL | 21µL | 30µL |
1 cut | 1.3µL | 0.2µL | -- | 1.0µL | 7.5µL | 10µL |
1 cut | 1.3µL | -- | 0.2µL | 1.0µL | 7.5µL | 10µL |
NC | 1.3µL | -- | -- | 1.0µL | 7.7µL | 10µL |
8/15 B0034(RBS) -(2) | SpeI | PstI | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|
2 cuts | 6.0µL | 1.0µL | 1.0µL | 3.0µL | 19µL | 30µL |
1 cut | 2.0µL | 0.2µL | -- | 1.0µL | 6.8µL | 10µL |
1 cut | 2.0µL | -- | 0.2µL | 1.0µL | 6.8µL | 10µL |
NC | 2.0µL | -- | -- | 1.0µL | 7.0µL | 10µL |
8/17 K117000(lysis1) -(1) | XbaI | PstI | BSA | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.0µL | 1.0µL | 1.0µL | 3.0µL | 3.0µL | 20µL | 30µL |
1 cut | 1.0µL | 0.2µL | -- | 1.0µL | 1.0µL | 6.8µL | 10µL |
1 cut | 1.0µL | -- | 0.2µL | 1.0µL | 1.0µL | 6.8µL | 10µL |
NC | 1.0µL | -- | -- | 1.0µL | 1.0µL | 7.0µL | 10µL |
8/17 K748002(lysis3) -(1) | XbaI | PstI | BSA | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.0µL | 1.0µL | 1.0µL | 3.0µL | 3.0µL | 20µL | 30µL |
1 cut | 1.0µL | 0.2µL | -- | 1.0µL | 1.0µL | 6.8µL | 10µL |
1 cut | 1.0µL | -- | 0.2µL | 1.0µL | 1.0µL | 6.8µL | 10µL |
NC | 1.0µL | -- | -- | 1.0µL | 1.0µL | 7.0µL | 10µL |
8/17 I13504(RBS-GFP-DT) -(1) | XbaI | PstI | BSA | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.0µL | 1.0µL | 1.0µL | 3.0µL | 3.0µL | 20µL | 30µL |
1 cut | 1.0µL | 0.2µL | -- | 1.0µL | 1.0µL | 6.8µL | 10µL |
1 cut | 1.0µL | -- | 0.2µL | 1.0µL | 1.0µL | 6.8µL | 10µL |
NC | 1.0µL | -- | -- | 1.0µL | 1.0µL | 7.0µL | 10µL |
8/17 I0462(RBS-luxR-DT) -(1) | XbaI | PstI | BSA | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.0µL | 1.0µL | 1.0µL | 3.0µL | 3.0µL | 20µL | 30µL |
1 cut | 1.0µL | 0.2µL | -- | 1.0µL | 1.0µL | 6.8µL | 10µL |
1 cut | 1.0µL | -- | 0.2µL | 1.0µL | 1.0µL | 6.8µL | 10µL |
NC | 1.0µL | -- | -- | 1.0µL | 1.0µL | 7.0µL | 10µL |
8/17 I732020(RBS-lacZα-DT) -(1) | XbaI | PstI | BSA | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|---|
2 cuts | 2.0µL | 1.0µL | 1.0µL | 3.0µL | 3.0µL | 20µL | 30µL |
1 cut | 1.0µL | 0.2µL | -- | 1.0µL | 1.0µL | 6.8µL | 10µL |
1 cut | 1.0µL | -- | 0.2µL | 1.0µL | 1.0µL | 6.8µL | 10µL |
NC | 1.0µL | -- | -- | 1.0µL | 1.0µL | 7.0µL | 10µL |
8/11 R6061 -(1) | SpeI | PstI | Buffer | MilliQ | total | |
---|---|---|---|---|---|---|
2 cuts | 8.0µL | 1.0µL | 1.0µL | 3.0µL | 17µL | 30µL |
1 cut | 3.0µL | 0.2µL | -- | 1.0µL | 5.8µL | 10µL |
1 cut | 3.0µL | -- | 0.2µL | 1.0µL | 5.8µL | 10µL |
NC | 3.0µL | -- | -- | 1.0µL | 6.0µL | 10µL |
- incubate at 37 °C, for 1 h
Electrophoresis
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1 | J23100 -② | SpeI | PstI |
2 | J23100 -② | SpeI | -- |
3 | J23100 -② | -- | PstI |
4 | J23100 -② | -- | -- |
5 | B0034(RBS) -(2) | SpeI | PstI |
6 | B0034(RBS) -(2) | SpeI | -- |
7 | B0034(RBS) -(2) | -- | PstI |
8 | B0034(RBS) -(2) | -- | -- |
9 | 1kb ladder | -- | -- |
10 | K11700(lysis1) -(1) | XbaI | PstI |
11 | K11700(lysis1) -(1) | XbaI | -- |
12 | K11700(lysis1) -(1) | -- | PstI |
13 | K11700(lysis1) -(1) | -- | -- |
14 | K748002(lysis3) -(1) | XbaI | PstI |
15 | K748002(lysis3) -(1) | XbaI | -- |
16 | K748002(lysis3) -(1) | -- | PstI |
17 | K748002(lysis3) -(1) | -- | -- |
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
18 | I13504(RBS-GFP-DT) -(1) | XbaI | PstI |
19 | I13504(RBS-GFP-DT) -(1) | XbaI | -- |
20 | I13504(RBS-GFP-DT) -(1) | -- | PstI |
21 | I13504(RBS-GFP-DT) -(1) | -- | -- |
22 | I0462(RBS-luxR-DT) -(1) | XbaI | PstI |
23 | I0462(RBS-luxR-DT) -(1) | XbaI | -- |
24 | I0462(RBS-luxR-DT) -(1) | -- | PstI |
25 | I0462(RBS-luxR-DT) -(1) | -- | -- |
26 | 1kb ladder | -- | -- |
27 | I732020(RBS-lacZα-DT) -(1) | XbaI | PstI |
28 | I732020(RBS-lacZα-DT) -(1) | XbaI | -- |
29 | I732020(RBS-lacZα-DT) -(1) | -- | PstI |
30 | I732020(RBS-lacZα-DT) -(1) | -- | -- |
31 | R0061(Plux) -(1) | SpeI | PstI |
32 | R0061(Plux) -(1) | SpeI | -- |
33 | R0061(Plux) -(1) | -- | PstI |
34 | R0061(Plux) -(1) | -- | -- |
Miniprep
DNA | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
8/17 PBR322(TaKaRa) -(1) | 47 | 1.68 | 1.55 |
8/17 K112806(lysis2) -(1) | 64 | 1.67 | 1.70 |