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Template:Kyoto/Notebook/Aug 17
From 2013.igem.org
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<span class="author">Murata, Murakami, Nakamoto, Ashida, Matsumoto and Yoshida</span> | <span class="author">Murata, Murakami, Nakamoto, Ashida, Matsumoto and Yoshida</span> | ||
{| class="wikitable" | {| class="wikitable" | ||
| - | ! ||8/11 J23100 -(2)|| | + | ! ||8/11 J23100 -(2)||SpeI||PstI||Buffer||MilliQ||total |
|- | |- | ||
|2 cuts||4.0µL||1.0µL||1.0µL||3.0µL||21µL||30µL | |2 cuts||4.0µL||1.0µL||1.0µL||3.0µL||21µL||30µL | ||
| Line 107: | Line 107: | ||
*at 37°C, for 1h | *at 37°C, for 1h | ||
{| class="wikitable" | {| class="wikitable" | ||
| - | ! ||8/15 B0034(RBS) -(2)|| | + | ! ||8/15 B0034(RBS) -(2)||SpeI||PstI||Buffer||MilliQ||total |
|- | |- | ||
|2 cuts||6.0µL||1.0µL||1.0µL||3.0µL||19µL||30µL | |2 cuts||6.0µL||1.0µL||1.0µL||3.0µL||19µL||30µL | ||
| Line 179: | Line 179: | ||
*at 37°C, for 1h | *at 37°C, for 1h | ||
{| class="wikitable" | {| class="wikitable" | ||
| - | ! ||8/11 R6061 -(1)|| | + | ! ||8/11 R6061 -(1)||SpeI||PstI||Buffer||MilliQ||total |
|- | |- | ||
|2 cuts||8.0µL||1.0µL||1.0µL||3.0µL||17µL||30µL | |2 cuts||8.0µL||1.0µL||1.0µL||3.0µL||17µL||30µL | ||
Revision as of 05:54, 24 September 2013
Contents |
Aug 17
Colony PCR
| Sample | base pair |
|---|---|
| 8/15 BBa_K112806(lysis2) -(1) | 828 |
| negative control | -- |
| 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 |
| negative control | -- |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 30s | 30cycles |
Liquid Culture
| Sample | medium |
|---|---|
| 8/15 BBa_K112806(lysis2) -(1) | Plusgrow medium(+CP) |
| 8/15 BBa_Q04400(tetR) -(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 |
- at 37°C, for 1h
| 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 |
- at 37°C, for 1h
| 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 |
- at 37°C, for 1h
| 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 |
- at 37°C, for 1h
| 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 |
- at 37°C, for 1h
| 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 |
- at 37°C, for 1h
| 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 |
- at 37°C, for 1h
| 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 |
- at 37°C, for 1h
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 |
