From 2013.igem.org
""m&m""
template
transformation
じっけんしゃ
| 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 | せいげんこーそ
|
| いかどうよう | | | | |
|
| | | | | |
|
Turing Pattern
Aug 20
Miniprep
by kojima
| 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
by Okazaki
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
by Kojima and Ashida
| 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
by Nakamoto
| 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℃ overnight 20:34~
Electrophoresis
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