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Revision as of 15:33, 20 August 2015
MARCH 2015
APRIL 2015
MAY 2015
JUNE 2015
JULY 2015
AUGUST 2015
SEPT. 2015
PCR pDawn BioBricks
PCR pDawn III
Aim: first step for pDawn amplification and mutagenesis
| pDawn III | |
|---|---|
| MQ water | 39,5 µL |
| Mg2+ 50 mM | 1,5 µL |
| Buffer RB | 5 µL |
| dNTPs 10µL | 1 µL |
| Primer Fwd 50µM | 1µL pDawn Fwd III |
| Primer Rev 50µM | 1µL pDawn Rev III |
| DNA | 1µL pDawn |
| Taq Pol | 0.5 µL |
| T°C | Time | Cycle | |
|---|---|---|---|
| Initial denaturation | |||
| Denaturation | |||
| Annealing | |||
| Extension | |||
| Final extension | |||
| Hold |
Gibson Assembly
Aim: first step of our VVD Biobrick
| Fragments | Concentration (ng.µL-1) | Lenght | Molecular weight (g.mol-1) | Molarity (mol.L-1) | 1/ratio | Dilution factor | Molarity after dilution | DNA Volume | Volume Water | Volume totale |
|---|---|---|---|---|---|---|---|---|---|---|
| pSB1C3 | |
0.000000063712 | 0.000000047103 | |||||||
| VVD1 | |
0.000000096698 | 0.000000047103 | |||||||
| VVD2 | |
0.000000047103 | 0.000000047103 | |||||||
| YC155 | |
0.000000493488 | 0.000000047103 |
DNA mix:
- YC155: 1µL + 9µL of MQ water
- VVD1: 1µL + 1µL of MQ water
- VVD2: no dilution
- pSB1C3: 3.5µL + 1.5µL of MQ water
5µl used for the Gibson assembly added to 15µl of master mix
Electrophoresis
Gel : 1,2 g agarose + 120 mL of TAE 1x + 150 µL of BET
Migration of pDawn III
Expected results
Results
Soft band at 1500 bp, good but not enough
1 July 15
Preparation of media
Aim: prepare media for bacteria culture
| LB Broth | LB Agar |
|---|---|
| 2.5g tryptone | 2.5g tryptone |
| 2.5g NaCl | 2.5g NaCl |
| 1.25g yeast extract | 1.25g yeast extract |
| 5.0g agar |
Qsp 250 ml distilled water
2 bottles of LB broth and 2 bottles of LB agar (+magnetic stirrer)
Auctoclave 121°C
28 March 15
Project
Our reality game using bacteria
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OPTOGENETICS
Our scientific core of the project
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MICROFLUIDICS
Our challenge in achieving the Bio-console
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