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Template:Kyoto/Notebook/Sep 6
From 2013.igem.org
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===Miniprep=== | ===Miniprep=== | ||
<div class="experiment"> | <div class="experiment"> | ||
| - | <span class="author"> | + | <span class="author">Honda and Kojima</span> |
{|class="wikitable" | {|class="wikitable" | ||
!DNA||concentration[µg/mL]||260/280||260/230 | !DNA||concentration[µg/mL]||260/280||260/230 | ||
|- | |- | ||
| - | |( | + | |PT181 antisense-(1)||238.7 ||1.68 ||1.44 |
| + | |- | ||
| + | |PT181 antisense-(2)||248.8 ||1.83 ||1.77 | ||
| + | |- | ||
| + | |PT181 antisense-(3)||223.2 ||1.78 ||1.61 | ||
| + | |- | ||
| + | |PT181 antisense-(4)||220.8 ||1.89 ||1.92 | ||
| + | |- | ||
| + | |tetR aptamer-(1)||252.5 ||1.89 ||1.90 | ||
| + | |- | ||
| + | |tetR aptamer-(2)||211.2 ||1.90 ||1.99 | ||
| + | |- | ||
| + | |PT181 attenuater||102.2 ||1.98 ||2.17 | ||
| + | |- | ||
| + | |121R aptamer-DT||251.7 ||1.92 ||2.01 | ||
| + | |- | ||
| + | |PSB4K5||212.9 ||1.80 ||1.90 | ||
| + | |- | ||
| + | |RBS-lacZα-DT||185.7 ||1.78 ||1.79 | ||
| + | |- | ||
| + | |RBS-lysis2-DT-(1)||84.8 ||1.85 ||2.24 | ||
|} | |} | ||
</div> | </div> | ||
Revision as of 08:02, 24 September 2013
Contents |
Sep 6
Liquid Culture
| Sample | medium |
|---|---|
| 9/4 RBS-lysis3＋DT3 | Plusgrow medium(+CP) |
| 9/4 RBS-lysis1＋DT3 | Plusgrow medium(+CP) |
| 9/4 psB4K5 | Plusgrow medium(+K) |
Colony PCR
| Sample | base pair |
|---|---|
| Pconst-spinach-DT-(1) | - |
| Pconst-spinach-DT-(2) | - |
| Pconst-spinach-DT-(3) | - |
| Pconst-spinach-DT-(4) | - |
| Plac-spinach-DT-(1) | - |
| Plac-spinach-DT-(2) | - |
| Plac-spinach-DT-(3) | - |
| Plac-spinach-DT-(4) | - |
| Pcon-PT181-antisense-(1) | - |
| Pcon-PT181-antisense-(2) | - |
| Pcon-PT181-antisense-(3) | - |
| Pcon-PT181-antisense-(4) | - |
| Sample | base pair |
|---|---|
| Ptet-PT181-antisense-(1) | - |
| Ptet-PT181-antisense-(2) | - |
| Ptet-PT181-antisense-(3) | - |
| Ptet-PT181-antisense-(4) | - |
| Plac-PT181-antisense-(1) | - |
| Plac-PT181-antisense-(2) | - |
| Plac-PT181-antisense-(3) | - |
| Plac-PT181-antisense-(4) | - |
Miniprep
| DNA | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| PT181 antisense-(1) | 238.7 | 1.68 | 1.44 |
| PT181 antisense-(2) | 248.8 | 1.83 | 1.77 |
| PT181 antisense-(3) | 223.2 | 1.78 | 1.61 |
| PT181 antisense-(4) | 220.8 | 1.89 | 1.92 |
| tetR aptamer-(1) | 252.5 | 1.89 | 1.90 |
| tetR aptamer-(2) | 211.2 | 1.90 | 1.99 |
| PT181 attenuater | 102.2 | 1.98 | 2.17 |
| 121R aptamer-DT | 251.7 | 1.92 | 2.01 |
| PSB4K5 | 212.9 | 1.80 | 1.90 |
| RBS-lacZα-DT | 185.7 | 1.78 | 1.79 |
| RBS-lysis2-DT-(1) | 84.8 | 1.85 | 2.24 |
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 |
