Notebook
Day 1
Description of everything we do
Voici le titre du tableau
| Titre A1 |
Titre A2 |
Titre A3 |
Titre A4 |
| Titre B1 |
Valeur B2 |
Valeur B3 |
Valeur B4 |
02/06/2015
- Stock solution MgSO4 : 0,5M
Pour 500mL :
61,62g de MgSO4 (MM : 246,48)
qsp 500mL H2O stérile
- SOB Medium : 1 L
| Final [c] |
Composent |
Volume & Mass |
| 2% |
Bactotryptone |
20 g |
| 10 mM |
NaCl |
2 mL (5 M stock) |
| 2% |
Yeast Extract |
5 g |
| 2,5 mM |
KCl |
0,18 g |
| 10 mM |
MgCl2 |
5 mL (2 M stock) |
| 10 mM |
MgSO4 |
20 mL (0,5 M) |
| - |
H2O |
qsp 1L |
| pH media to 7 with NaOH, then autoclave |
- 2XTY Medium 500 mL
| Composent |
Volume & Mass |
| Bactotryptone |
8 g |
| NaCl |
2,5 g |
| Yeast Extract |
5 g |
| H2O |
qsp 500 mL |
03/06/2015
- Transformation Broth (TB): 1 L
| Final [c] |
Composent |
Volume & Mass |
| 10 mM |
Pipes |
3,02 g |
| 15 mM |
CaCl2 |
2,21 g |
| 250 mM |
KCl |
18,64 g |
Mix the Pipes, CaCl2, and KCl in 900 ml of millipore water. Add NaOH until pH is 6.7,
Don't worry, dust disappear after pH adjust.
Add MnCl2 (see below), stir, adjust volume to 1 L, then filter sterilize. Store at 4C.
| Final [c] |
Composent |
Volume & Mass |
| 55 mM |
MnCl2 |
10,89 g |
- Making DMSO Competent Cells : DAY ONE
Grow 12 ml overnight culture of favorite strain of E. coli in 2XTY at 37°C and 250 rpm (We use Denis’ DH5-alpha).