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Template:Kyoto/Notebook/Sep 1
From 2013.igem.org
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===M9 Medium=== | ===M9 Medium=== | ||
Revision as of 00:39, 23 September 2013
Contents |
Sep 1
Transformation
| Name | Sample | Competent Cells | Total | Plate |
|---|---|---|---|---|
| 8/31 tRNA-Spinach(pSB1C3) | 2µL | 20µL | 22µL | CP |
| 8/31 pT181 antisense(pSB1C3) | 2µL | 20µL | 22µL | CP |
| 8/31 pT181 attenuator(pSB1C3) | 2µL | 20µL | 22µL | CP |
| 8/31 pT181 antisense(pSB1C3) | 2µL | 20µL | 22µL | CP |
| 8/31 pT181 attenuator(2)-DT | 2µL | 20µL | 22µL | CP |
| 8/31 pT181 antisense-DT | 2µL | 20µL | 22µL | CP |
| 8/31 tRNA-Spniach-DT | 2µL | 20µL | 22µL | CP |
| 8/31 RBS-lysis2-DT | 2µL | 20µL | 22µL | CP |
| 8/31 Pconst-RBS-tetR-DT | 2µL | 20µL | 22µL | Amp |
| 8/31 Plac-RBS-lacZα-DT | 2µL | 20µL | 22µL | CP |
| 8/31 Ptet-RBS-lacZα-DT | 2µL | 20µL | 22µL | CP |
| 8/31 | 2µL | 20µL | 22µL | CP |
ligation production PCR
| Sample | base pair |
|---|---|
| 8/31 tRNA Spinach(pSB1C3) | 459 |
| 8/31 pT181 antisense(2)(pSB1C3)(E+S) | 415 |
| 8/31 pT181 attenuator(2)(pSB1C3)(X+P) | 601 |
| 8/31 pT181 antisense(2)(pSB1C3)(X+P) | 415 |
| 8/31 pT181 attenuator(2)-DT | 754 |
| 8/31 pT181 antisense(2)-DT | 577 |
| 8/31 tRNA Spinach-DT | 596 |
| 8/22 pSB1C3(2)(controll) | 1383 |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 30min | 30cycle |
Ligation
Liquid Culture
M9 Medium
5xM9 liquid medium
| volume | 100mL |
|---|---|
| NaHPO4 | 30g |
| KH2PO | 3g |
| Nacl | 0.25g |
| NH4Cl | 0.5g |
| 3.75% agar solution | 400mL |
| 1M MgSO4 | 0.5mL |
| 2M Glucose | 2.8mL |
| 1% Vitamin B1 | 0.5mL |
| 1M CaCl2 | 0.05mL |
1. mix 5xM9 medium and 3.75%Agar solution 400mL
2. Add 1M MgSO4, 2M Glucose and 1 VitaminB1 0.5mL
3. Add CaCl2 while shaking Erlenmeyer flask.
