Template:Kyoto/Notebook/Aug 17
From 2013.igem.org
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</div> | </div> | ||
+ | |||
+ | ===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 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 | ||
+ | </div> | ||
+ | |||
===Master Plate=== | ===Master Plate=== | ||
<div class="experiment"> | <div class="experiment"> |
Revision as of 15:49, 23 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) |
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 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
Master Plate
Number | Use LB plate (+CP) |
---|---|
1 | BBa_K112806(lysis2) -(1) |
Number | Use LB plate (+Kana) |
---|---|
1 | BBa_Q04400(tetR) -(1) |