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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"> | ||
| Line 124: | Line 125: | ||
|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 | ||
| Line 172: | Line 169: | ||
|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 | ||
| Line 184: | Line 180: | ||
|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 | ||
| Line 196: | Line 191: | ||
|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 | ||
| Line 208: | Line 202: | ||
|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> | ||
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
| Lane | Sample |
|---|---|
| 1 | 1kbp ladder |
| 2 | K112806(lysis2) -1 |
| 3 | NC |
| 4 | Q04400(tetR) -1 |
| 5 | NC |
| 6 | 1kbp ladder |
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) | -- | -- |
.jpg)
| 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) | -- | -- |
.jpg)
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 |
