"
Template:Kyoto/Notebook/Sep 23
From 2013.igem.org
(Difference between revisions)
(→Sep 23) |
(→Ligation) |
||
| Line 62: | Line 62: | ||
<!-- こっから --> | <!-- こっから --> | ||
<div class="experiment"> | <div class="experiment"> | ||
| - | <span class="author"> | + | <span class="author">Yoshida</span> |
{| class="wikitable" | {| class="wikitable" | ||
!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | !state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | ||
|- | |- | ||
| - | |experiment||9/21 Pcon-RBS-GFP-DT E+X||47.5 µg/mL||9/15 Pcon-RBS-tetR-DT E+S|| | + | |experiment||9/21 Pcon-RBS-GFP-DT E+X||47.5 µg/mL||9/15 Pcon-RBS-tetR-DT E+S||59 µg/mL||2.3 µg/mL |
|- | |- | ||
| - | |experiment||9/21 Pcon-RBS-GFP-DT E+X||47.5 µg/mL||9/15 Pcon-apt12_1R-DT E+S|| | + | |experiment||9/21 Pcon-RBS-GFP-DT E+X||47.5 µg/mL||9/15 Pcon-apt12_1R-DT E+S||17 µg/mL||2.4 µg/mL |
|- | |- | ||
| - | |experiment||9/21 Pcon-RBS-GFRDT E+X|| µg/mL||9/15 Pcon-DT181 atte-DT|| | + | |experiment||9/21 Pcon-RBS-GFRDT E+X||2.1 µg/mL||9/15 Pcon-DT181 atte-DT||21 µg/mL||3.0 µg/mL |
|- | |- | ||
| - | |experiment||9/22 apt 12_1R-DT E+X||67.8 µg/mL||9/18 Pcon-DT181 atte E+S|| | + | |experiment||9/22 apt 12_1R-DT E+X||67.8 µg/mL||9/18 Pcon-DT181 atte E+S||5.7 µg/mL||3.5 µg/mL |
|- | |- | ||
| - | |experiment||9/22RBS-GFP-DT E+X||87.7 µg/mL|| | + | |experiment||9/22RBS-GFP-DT E+X||87.7 µg/mL||9/18 Pcon-DT18 atte E+S||6.7 µg/mL||3.5 µg/mL |
|- | |- | ||
| - | |experiment||9/13 DT E+X||25.4 µg/mL|| | + | |experiment||9/13 DT E+X||25.4 µg/mL||9/22 Pcon-DT181 anti E+S||46.8 µg/mL||2.3 µg/mL |
|} | |} | ||
</div> | </div> | ||
<!-- ここまでをコピーしてね --> | <!-- ここまでをコピーしてね --> | ||
Revision as of 04:24, 24 September 2013
Contents |
Sep 23
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 1kbp ladder | |
| 3-4 | Pcon-RBS-tetR-PT |
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| Pcon-PT181 anti E+S | 46.7 | 1.06 | 0.93 |
| Pcon-RBS-tetR-DT E+X | 119.3 | 0.99 | |
| RBS-8FP-DT E+X | 87.7 | 1.00 | 0.98 |
| ap42_1R-DT E+X | 68.8 | 1.32 | 1.03 |
| Pcon-RBS-tetR-DT 1-2 A | 203.0 | 1.66 | 1.79 |
| Pcon-RBS-tetR-DT 3-2 A | 67.7 | 2.64 | 1.28 |
| Pcon-RBS-GFP-DT 1-S A | 185.6 | 1.88 | 2.04 |
| Pcon-RBS-GFP-DT 2-S A | 298.6 | 1.68 | 1.55 |
| Pcon-tetR-DT E-2 A | 202.4 | 1.57 | 1.66 |
PCR
| genome DNA | KOD plus | 10x buffer | dNTP | MgSO4 | SasA_fwd primer | SasA_rev primer | MilliQ | total |
|---|---|---|---|---|---|---|---|---|
| 9/8 Pcon-pT181 anti | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 15.5 | 25 |
| 8/20 Ptet(1) | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 15.5 | 25 |
| 9/16 Plac 2 | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 15.5 | 25 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94 °C | 98 °C | 57 °C | 68 °C | -- |
| 2min | 10sec | 30sec | 20sec | 30cycles |
Ligation
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | 9/21 Pcon-RBS-GFP-DT E+X | 47.5 µg/mL | 9/15 Pcon-RBS-tetR-DT E+S | 59 µg/mL | 2.3 µg/mL |
| experiment | 9/21 Pcon-RBS-GFP-DT E+X | 47.5 µg/mL | 9/15 Pcon-apt12_1R-DT E+S | 17 µg/mL | 2.4 µg/mL |
| experiment | 9/21 Pcon-RBS-GFRDT E+X | 2.1 µg/mL | 9/15 Pcon-DT181 atte-DT | 21 µg/mL | 3.0 µg/mL |
| experiment | 9/22 apt 12_1R-DT E+X | 67.8 µg/mL | 9/18 Pcon-DT181 atte E+S | 5.7 µg/mL | 3.5 µg/mL |
| experiment | 9/22RBS-GFP-DT E+X | 87.7 µg/mL | 9/18 Pcon-DT18 atte E+S | 6.7 µg/mL | 3.5 µg/mL |
| experiment | 9/13 DT E+X | 25.4 µg/mL | 9/22 Pcon-DT181 anti E+S | 46.8 µg/mL | 2.3 µg/mL |
