Template:Kyoto/Notebook/Sep 6
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==Sep 6== | ==Sep 6== | ||
+ | ===Colony PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !|Sample||base pair | ||
+ | |- | ||
+ | |8/15 BBa_K112806(lysis2) -(1)||828 | ||
+ | |- | ||
+ | |negative control||-- | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |5min||30s||30s||1min||30cycles | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |8/10 BBa_Q0400 -(1)||3995 | ||
+ | |- | ||
+ | |negative control||-- | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |5min||30s||30s||30s||30cycles | ||
+ | |} | ||
+ | |||
+ | ===Liquid Culture=== | ||
+ | <!-- ここから --> | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||medium | ||
+ | |- | ||
+ | |(・ω・) || | ||
+ | |- | ||
+ | |(・X・) || | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | |||
+ | ===Colony PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !|Sample||base pair | ||
+ | |- | ||
+ | |8/15 BBa_K112806(lysis2) -(1)||828 | ||
+ | |- | ||
+ | |negative control||-- | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |5min||30s||30s||1min||30cycles | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |8/10 BBa_Q0400 -(1)||3995 | ||
+ | |- | ||
+ | |negative control||-- | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |5min||30s||30s||30s||30cycles | ||
+ | |} | ||
+ | |||
+ | ===Miniprep=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">No name</span> | ||
+ | {|class="wikitable" | ||
+ | !DNA||concentration[µg/mL]||260/280||260/230 | ||
+ | |- | ||
+ | |(・▽・)|| || || | ||
+ | |} | ||
+ | </div> | ||
+ | <!-- ここまでをコピーしてね --> | ||
+ | |||
+ | ===Restriction Enzyme Digestion=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Murata, Murakami, Nakamoto, Ashida, Matsumoto and Yoshida</span> | ||
+ | {| class="wikitable" | ||
+ | ! ||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 | ||
+ | {| class="wikitable" | ||
+ | ! ||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 | ||
+ | {| class="wikitable" | ||
+ | ! ||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 | ||
+ | {| class="wikitable" | ||
+ | ! ||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 | ||
+ | {| class="wikitable" | ||
+ | ! ||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 | ||
+ | {| class="wikitable" | ||
+ | ! ||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 | ||
+ | {| class="wikitable" | ||
+ | ! ||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 | ||
+ | {| class="wikitable" | ||
+ | ! ||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 | ||
+ | </div> | ||
+ | |||
+ | ===Gel Extraction=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Ashida</span> | ||
+ | {| class="wikitable" | ||
+ | !Lane||DNA||Enzyme | ||
+ | |- | ||
+ | |1||100bp ladder||-- | ||
+ | |- | ||
+ | |3||8/18 K117000(lysis2) -1||XbaI & PstI | ||
+ | |- | ||
+ | |5||8/18 K117000(lysis2) -1||XbaI & PstI | ||
+ | |} | ||
+ | [[File:igku_Aug19electrophoresis3]]<br> | ||
+ | [[File:igku_Aug19electrophoresis4]]<br> | ||
+ | {| class="wikitable" | ||
+ | !Name||concentration[µg/mL]||260/280||260/230 | ||
+ | |- | ||
+ | |8/18 lysis2-1(XbaI & PstI)||3.1||3.64||0.19 | ||
+ | |- | ||
+ | |8/18 lysis2-1(XbaI & PstI)||3.4||1.89||0.13 | ||
+ | |} | ||
+ | |||
+ | </div> | ||
+ | |||
+ | |||
+ | ===Liquid Culture=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !Sample||medium | ||
+ | |- | ||
+ | |8/15 BBa_K112806(lysis2) -(1)||Plusgrow medium(+CP) | ||
+ | |- | ||
+ | |8/15 BBa_Q04400(tetR) -(1)||Plusgrow medium(+Amp) | ||
+ | |- | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | |||
+ | ===Colony PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !|Sample||base pair | ||
+ | |- | ||
+ | |8/15 BBa_K112806(lysis2) -(1)||828 | ||
+ | |- | ||
+ | |negative control||-- | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |5min||30s||30s||1min||30cycles | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |8/10 BBa_Q0400 -(1)||3995 | ||
+ | |- | ||
+ | |negative control||-- | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |5min||30s||30s||30s||30cycles | ||
+ | |} | ||
+ | |||
+ | ===Colony PCR=== | ||
+ | <div class="experiment"> | ||
+ | <span class="author">Nakamoto</span> | ||
+ | {| class="wikitable" | ||
+ | !|Sample||base pair | ||
+ | |- | ||
+ | |8/15 BBa_K112806(lysis2) -(1)||828 | ||
+ | |- | ||
+ | |negative control||-- | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |5min||30s||30s||1min||30cycles | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !Sample||base pair | ||
+ | |- | ||
+ | |8/10 BBa_Q0400 -(1)||3995 | ||
+ | |- | ||
+ | |negative control||-- | ||
+ | |} | ||
+ | {| class="wikitable" | ||
+ | !PreDenature||Denature||Annealing||Extension||cycle | ||
+ | |- | ||
+ | |94°C||94°C||55°C||68°C||-- | ||
+ | |- | ||
+ | |5min||30s||30s||30s||30cycles | ||
+ | |} |
Revision as of 07:24, 24 September 2013
Contents |
Sep 6
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 |
---|---|
(・ω・) | |
(・X・) |
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 |
Miniprep
DNA | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
(・▽・) |
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
Gel Extraction
Lane | DNA | Enzyme |
---|---|---|
1 | 100bp ladder | -- |
3 | 8/18 K117000(lysis2) -1 | XbaI & PstI |
5 | 8/18 K117000(lysis2) -1 | XbaI & PstI |
File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4
Name | concentration[µg/mL] | 260/280 | 260/230 |
---|---|---|---|
8/18 lysis2-1(XbaI & PstI) | 3.1 | 3.64 | 0.19 |
8/18 lysis2-1(XbaI & PstI) | 3.4 | 1.89 | 0.13 |
Liquid Culture
Sample | medium |
---|---|
8/15 BBa_K112806(lysis2) -(1) | Plusgrow medium(+CP) |
8/15 BBa_Q04400(tetR) -(1) | Plusgrow medium(+Amp) |
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 |
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 |