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Template:Kyoto/Notebook/Sep 8
From 2013.igem.org
(Difference between revisions)
(→Gel Extraction) |
(→Restriction Enzyme Digestion) |
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<span class="author">じっけんしゃ</span> | <span class="author">じっけんしゃ</span> | ||
{| class="wikitable" | {| class="wikitable" | ||
| - | ! || | + | ! ||9/8 Pcon-pT181 anti||E||S||X||P||BSA||Buffer||MilliQ||total |
|- | |- | ||
| - | |2 cuts|| | + | |2 cuts||8.2µL|| 0µL|| 1µL|| 0µL|| 1µL|| 3µL|| 3µL|| 13.8µL|| 30µL |
|- | |- | ||
| - | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL | + | |NC|| 0.4µL|| 0µL|| 0µL|| 0µL|| 0µL|| 1µL|| 1µL|| 7.6µL|| 10µL |
|} | |} | ||
| - | |||
| - | |||
{| class="wikitable" | {| class="wikitable" | ||
| - | ! || | + | ! ||9/8 Pcon-spin-DT||E||S||X||P||BSA||Buffer||MilliQ||total |
|- | |- | ||
| - | |2 cuts|| | + | |2 cuts||7.9µL|| 1µL|| 1µL|| 0µL|| 0µL|| 3µL|| 3µL||14.1µL|| 30µL |
|- | |- | ||
| - | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL | + | |NC|| 0.4µL|| 0µL|| 0µL|| 0µL|| 0µL|| 1µL|| 1µL||7.6µL|| 10µL |
|} | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/21 F1 atte||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts|| 6.5µL|| 0µL|| 0µL|| 1µL|| 1µL|| 3µL|| 3µL||15.5µL||30µL | ||
| + | |- | ||
| + | |NC|| 0.3µL||0µL||0µL|| 0µL|| 0µL||1µL||1µL||7.7µL||10µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/21 F3m2 atte||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/21 F1 anti||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/21 F6 anti||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/21 tetR aptamer 12_1M||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/21 tetR aptamer 12_P||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||9/4 spin-DT||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||9/7 RBS-lysis1-DT||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||9/6 RBS-lysis2-DT||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||9/7 RBS-lysis3-DT||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/20 J23100 RBS-lacZα-DT-(1)||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | {| class="wikitable" | ||
| + | ! ||8/20 RBS-lacZα-DT||E||S||X||P||BSA||Buffer||MilliQ||total | ||
| + | |- | ||
| + | |2 cuts||µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |- | ||
| + | |NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL||µL|| µL | ||
| + | |} | ||
| + | |||
===Transformation=== | ===Transformation=== | ||
<!-- こっから --> | <!-- こっから --> | ||
Revision as of 00:51, 24 September 2013
Contents |
Sep 8
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | |
| 2 | RBS-lysis1-DT | XbaI & PstI |
| 3 | RBS-lysis1-DT | XbaI & PstI |
| 5 | RBS-lysis3-DT | XbaI & PstI |
| 6 | RBS-lysis3-DT | XbaI & PstI |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| RBS-lysis1-DT (XbaI & PstI) | 5.3 | 1.98 | 0.06 |
| RBS-lysis3-DT (XbaI & PstI) | 5.3 | 1.98 | 0.06 |
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100bp ladder | |
| 3 | Pcon-pT181 attenuator | EcoRI & SpeI |
| 4 | Pcon-pT181 attenuator | EcoRI & SpeI |
| 6 | Spinach-DT | XbaI & PstI |
| 7 | Spinach-DT | XbaI & PstI |
| 9 | RBS-lacZα-DT | XbaI & PstI |
| 10 | RBS-lacZα-DT | XbaI & PstI |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| Pcon-pT181 attenuator (EcoRI & SpeI) | 5.2 | 1.82 | 0.36 |
| Spinach-DT (XbaI & PstI) | 8.0 | 1.87 | 0.25 |
| RBS-lacZα-DT (XbaI & PstI) | 10.3 | 1.71 | 0.54 |
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 1kbp ladder | |
| 2 | Ptet-pT181 antisense | EcoRI & PstI |
| 3 | Ptet-pT181 antisense | EcoRI & PstI |
| 5 | pSB4K5 | EcoRI & SpeI |
| 6 | pSB4K5 | EcoRI & SpeI |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| Ptet-pT181 antisense(EcoRI & PstI) | 30.6 | 1.84 | 1.16 |
| pSB4K5 (EcoRI & SpeI) | 21.8 | 1.87 | 0.98 |
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 1kbp ladder | |
| 2 | Pcon-pT181 attenuator | SpeI & PstI |
| 3 | Pcon-pT181 attenuator | SpeI & PstI |
| 5 | Plac | SpeI & PstI |
| 6 | Plac | SpeI & PstI |
| 8 | pSB1C3 | EcoRI & SpeI |
| 9 | pSB1C3 | EcoRI & SpeI |
| 11 | pSB1C3 | XbaI & PstI |
| 12 | pSB1C3 | XbaI & PstI |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| Pcon-pT181 attenuator (SpeI & PstI) | 14.5 | 1.91 | 1.09 |
| Plac (SpeI & PstI) | 11.6 | 1.74 | 1.19 |
| pSB1C3 (EcoRI & SpeI) | 5.7 | 1.74 | 0.34 |
| pSB1C3 (XbaI & PstI) | 5.6 | 2.00 | 0.37 |
Miniprep
| DNA | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| Pcon-pT181 anti | 243.0 | 1.90 | 1.81 |
| Pcon-Spinach-DT | 253.7 | 1.57 | 1.51 |
Restriction Enzyme Digestion
| 9/8 Pcon-pT181 anti | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | 8.2µL | 0µL | 1µL | 0µL | 1µL | 3µL | 3µL | 13.8µL | 30µL |
| NC | 0.4µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL |
| 9/8 Pcon-spin-DT | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | 7.9µL | 1µL | 1µL | 0µL | 0µL | 3µL | 3µL | 14.1µL | 30µL |
| NC | 0.4µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.6µL | 10µL |
| 8/21 F1 atte | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | 6.5µL | 0µL | 0µL | 1µL | 1µL | 3µL | 3µL | 15.5µL | 30µL |
| NC | 0.3µL | 0µL | 0µL | 0µL | 0µL | 1µL | 1µL | 7.7µL | 10µL |
| 8/21 F3m2 atte | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 8/21 F1 anti | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 8/21 F6 anti | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 8/21 tetR aptamer 12_1M | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 8/21 tetR aptamer 12_P | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 9/4 spin-DT | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 9/7 RBS-lysis1-DT | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 9/6 RBS-lysis2-DT | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 9/7 RBS-lysis3-DT | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 8/20 J23100 RBS-lacZα-DT-(1) | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| 8/20 RBS-lacZα-DT | E | S | X | P | BSA | Buffer | MilliQ | total | |
|---|---|---|---|---|---|---|---|---|---|
| 2 cuts | µL | µL | µL | µL | µL | µL | µL | µL | µL |
| NC | µL | µL | µL | µL | µL | µL | µL | µL | µL |
Transformation
| Name | Sample | Competent Cells | Total | Plate |
|---|---|---|---|---|
| いでんし | なんちゃらµL | ごにょごにょµL | ほにゃららµL | こーせーぶっしつ |
| いかどうよう | ||||
Electrophoresis
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| （°Д°） |
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| （°Д°） |
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| （°Д°） |
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| （＞ω・）b | ||
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|---|
| （´ω｀） | ||||
| （ΦωΦ^） |
| Lane | DNA | Enzyme |
|---|---|---|
| （＞ω・）b | ||
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|---|
| （´ω｀） | ||||
| （ΦωΦ^） |
| Lane | DNA | Enzyme |
|---|---|---|
| （＞ω・）b | ||
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|---|
| （´ω｀） | ||||
| （ΦωΦ^） |
| Lane | DNA | Enzyme |
|---|---|---|
| （＞ω・）b | ||
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|---|
| （´ω｀） | ||||
| （ΦωΦ^） |
| Lane | DNA | Enzyme |
|---|---|---|
| （＞ω・）b | ||
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|---|
| （´ω｀） | ||||
| （ΦωΦ^） |
Ligation
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
| NC | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
| NC | name | xx µg/mL | name | xx µg/mL | xx µg/mL |
PCR
| genome DNA | KOD plus | 10x buffer | dNTP | MgSO4 | SasA_fwd primer | SasA_rev primer | MilliQ | total |
|---|---|---|---|---|---|---|---|---|
| 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 25 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| °C | °C | °C | °C | -- |
| （◎Д◎） |
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| （＞ω・）b | ||
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|---|
| （´ω｀） | ||||
| （ΦωΦ^） |
| Lane | DNA | Enzyme |
|---|---|---|
| （＞ω・）b | ||
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | quantity | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|---|
| （´ω｀） | ||||
| （ΦωΦ^） |
