Team:Kyoto/Notebook
From 2013.igem.org
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|Fusion6 antisense||1µL||10µL||11µL||8/19 LB+Amp | |Fusion6 antisense||1µL||10µL||11µL||8/19 LB+Amp | ||
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- | incubate | + | incubate 37°C overnight 20:34~ |
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- | incubate | + | incubate 37°C 10hour |
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<!-- 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 03:44, 23 August 2013
Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Material and method | Safety | Attributions |
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Contents |
template
transformation
Name | Well | Sample | Competent Cells | Total | Plate |
---|---|---|---|---|---|
いでんし | ぱーつのうぇる | 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 | |||||
Aug 20
Miniprep
DNA | concentration[µg/µL] | 260/280 | 260/230 |
---|---|---|---|
8/18 Ptet -1 | 220 | 1.69 | 1.87 |
8/18 Ptet -2 | 210 | 1.69 | 1.71 |
8/18 RBS-tetR-DT -1 | 236 | 1.63 | 1.69 |
8/18 RBS-tetR-DT -2 | 182 | 1.65 | 1.98 |
8/18 J23100-RBS-GFP-DT -1 | 334 | 1.71 | 2.12 |
8/18 J23100-RBS-luxR-DT -1 | 440 | 1.67 | 1.60 |
8/18 J23100-RBS-luxR-DT -2 | 344 | 1.68 | 1.83 |
8/18 J23100-RBS-lacZα-DT -1 | 280 | 1.70 | 1.81 |
8/18 RBS-lysis3 -1 | 282 | 1.67 | 1.76 |
8/18 RBS-lysis3 -2 | 212 | 1.30 | 1.35 |
LB Culture Medium
volume | 200mL |
---|---|
Bacto Trypton | 2g |
Bacto yeast extract | 1g |
Nacl | 1g |
Agar Powder | 2g |
Amp | 40µl |
1.Measuring reagents and put they in the Erlenmeyer flask
2.diluting in measuring cylinder to 200ml total
3.autoclaving
4.adding ampicirine 40ml
not written yet
Ristriction Enzyme Proccessing
8/20 Ptet -1(220µg/mL) | EcoRI | XbaI | 10x BSA | 10x buffer M | MilliQ | total |
---|---|---|---|---|---|---|
4.5 | 1 | 1 | 3 | 3 | 17.5 | 30 |
0.5 | 0.2 | 0 | 1 | 1 | 7.3 | 10 |
0.5 | 0 | 0.2 | 1 | 1 | 7.3 | 10 |
0.5 | 0 | 0 | 1 | 1 | 7.5 | 10 |
8/20 RBS-tetR-DT -2(182µg/mL) | EcoRI | SpeI | 10x buffer H | MilliQ | total |
---|---|---|---|---|---|
5.5 | 1 | 1 | 3 | 19.5 | 30 |
0.6 | 0.2 | 0 | 1 | 8.2 | 10 |
0.6 | 0 | 0.2 | 1 | 8.2 | 10 |
0.6 | 0 | 0 | 1 | 8.4 | 10 |
8/20 J23100-RBS-GFP-DT(334µg/ml) | EcoRI | SpeI | 10x buffer H | MilliQ | total |
---|---|---|---|---|---|
3.0 | 1 | 1 | 3 | 22 | 30 |
0.3 | 0.2 | 0 | 1 | 8.5 | 10 |
0.3 | 0 | 0.2 | 1 | 8.5 | 10 |
0.3 | 0 | 0 | 1 | 8.7 | 10 |
8/20 J23100-RBS-luxR-DT -2(344µg/mL) | EcoRI | XbaI | BSA | 10x buffer M | MilliQ | total |
---|---|---|---|---|---|---|
2.9 | 1 | 1 | 3 | 3 | 19.1 | 30 |
0.3 | 0.2 | 0 | 1 | 1 | 7.5 | 10 |
0.3 | 0 | 0.2 | 1 | 1 | 7.5 | 10 |
0.3 | 0 | 0 | 1 | 1 | 7.7 | 10 |
8/20 RBS-lysis3 -1(282µg/mL) | EcoRI | SpeI | 10x buffer H | MilliQ | total |
---|---|---|---|---|---|
3.5 | 1 | 1 | 3 | 21.5 | 30 |
0.4 | 0.2 | 0 | 1 | 8.4 | 10 |
0.4 | 0 | 0.2 | 1 | 8.4 | 10 |
0.4 | 0 | 0 | 1 | 8.6 | 10 |
8/17 DT -1(188µg/mL) | EcoRI | XbaI | 10x BSA | 10x buffer M | MilliQ | total |
---|---|---|---|---|---|---|
5.3 | 1 | 1 | 3 | 3 | 16.7 | 30 |
0.3 | 0.2 | 0 | 1 | 1 | 7.5 | 10 |
0.3 | 0 | 0.2 | 1 | 1 | 7.5 | 10 |
0.3 | 0 | 0 | 1 | 1 | 7.7 | 10 |
incubate ~20:00
Transformation
Name | Sample | Competent Cells(XL10-gold) | Total | Plate |
---|---|---|---|---|
tRNA-Spinach-tRNA | 1µL | 10µL | 11µL | 8/19 LB+Amp |
tetR-aptamer 12_PC(076) | 1µL | 10µL | 11µL | 8/19 LB+Amp |
tetR-aptamer 12_1R(113) | 1µL | 10µL | 11µL | 8/19 LB+Amp |
tetR-aptamer 12_1M(105) | 1µL | 10µL | 11µL | 8/19 LB+Amp |
pT181 attenuator | 1µL | 10µL | 11µL | 8/19 LB+Amp |
Fusin1 attenuator | 1µL | 10µL | 11µL | 8/19 LB+Amp |
Fusion3m2 attenuator | 1µL | 10µL | 11µL | 8/19 LB+Amp |
pT181 antisense | 1µL | 10µL | 11µL | 8/19 LB+Amp |
Fusion1 antisense | 1µL | 10µL | 11µL | 8/19 LB+Amp |
Fusion6 antisense | 1µL | 10µL | 11µL | 8/19 LB+Amp |
incubate 37°C overnight 20:34~
Electrophoresis
Lane | Sample | Enzyme1 | Enzyme2 |
---|---|---|---|
1 | 8/20 Ptet -1 | EcoRI | XbaI |
2 | 8/20 Ptet -1 | EcoRI | - |
3 | 8/20 Ptet -1 | - | XbaI |
4 | 8/20 Ptet -1 | - | - |
5 | 8/20 RBS-tetR-DT -2 | EcoRI | SpeI |
6 | 8/20 RBS-tetR-DT -2 | EcoRI | - |
7 | 8/20 RBS-tetR-DT -2 | - | SpeI |
8 | 8/20 RBS-tetR-DT -2 | - | - |
9 | 1kbp ladder | - | - |
10 | 8/20 J23100-RBS-GFP-DT -1 | EcoRI | SpeI |
11 | 8/20 J23100-RBS-GFP-DT -1 | EcoRI | - |
12 | 8/20 J23100-RBS-GFP-DT -1 | - | SpeI |
13 | 8/20 J23100-RBS-GFP-DT -1 | - | - |
14 | 8/20 J23100-RBS-luxR-DT -1 | EcoRI | XbaI |
15 | 8/20 J23100-RBS-luxR-DT -1 | EcoRI | - |
16 | 8/20 J23100-RBS-luxR-DT -1 | - | XbaI |
17 | 8/20 J23100-RBS-luxR-DT -1 | - | - |
18 | 1kbp ladder | - | - |
19 | 8/20 RBS-lysis3 -1 | EcoRI | SpeI |
20 | 8/20 RBS-lysis3 -1 | EcoRI | - |
21 | 8/20 RBS-lysis3 -1 | - | SpeI |
22 | 8/20 RBS-lysis3 -1 | - | - |
23 | 1kbp ladder | - | - |
24 | 8/17 DT | EcoRI | XbaI |
25 | 8/17 DT | EcoRI | - |
26 | 8/17 DT | - | XbaI |
27 | 8/17 DT | - | - |
Aug 21
Plusgrow medium(+amp)
- material
- 8/17 Plusgrow medium 50ml
- 7/22 5000× Ampicillin 10µl
- Measuring materials and putting them in a 50ml tube
Liquid culture
Sample | medium | |
---|---|---|
8/20 tRNA-spinach | 8/21 Plusgrow medium(+amp) | |
8/20 TetR-aptamer 12_P(1076) | 8/21 Plusgrow medium(+amp) | |
8/20 TetR-aptamer 12_1R(113) | 8/21 Plusgrow medium(+amp) | |
8/20 TetR-aptamer 12_1M(105) | 8/21 Plusgrow medium(+amp) | |
8/20 PT181 attenuator | 8/21 Plusgrow medium(+amp) | |
8/20 Fusion1 attenuator | 8/21 Plusgrow medium(+amp) | |
8/20 Fusion3m2 attenuator | 8/21 Plusgrow medium(+amp) | |
8/20 PT181 antisense | 8/21 Plusgrow medium(+amp) | |
8/20 Fusion1 antisense | 8/21 Plusgrow medium(+amp) | |
8/20 Fusion6 antisense | 8/21 Plusgrow medium(+amp) |
incubate 37°C 10hour