Difference between revisions of "Team:Grenoble-Alpes/Protocols"

Preparations

LB Broth

Materials

20g LB Broth Base (powder)

1L Distillated Water

Methods

1. Add 20g of LB Broth Base per 1L of distillated water
2. Homogenize
3. Autoclave at 121°C for 15 minutes

LB Agar

Materials

32g LB Agar (powder)

1L Distilled Water

Methods

1. Add 32g of LB Agar per 1L of distilled water
2. Homogenize
3. Autoclave at 121°C for 15 minutes

Chloramphenicol (Cam) 25ug/mL Stock

Materials

0,25g Chloramphenicol (25mg/mL)

10mL Ethanol

Methods

1. Add 0,25g of Cam per 10mL of ethanol
2. Homogenize by vortexing
3. Filter with a 22mm membrane filter
4. Conserve at -20°C in 1mL aliquot

Petri Dish Preparation

Materials

20mL LB Agar

20µL Cam (25µg/mL)

Methods

1. Add of 20uL Cam in 20mL of liquid LB Agar
2. Pour the solution in the plate
3. Wait until the agar solidifies
4. Conserve returned in 4°C

Glycerol 80%

Materials

80mL Glycerol 100%

20mL Sterile Water

Methods

1. Harvest 80mL of glycerol 100%
2. Add 20mL of sterile water
3. Conserve at 4°C

CaCl2 100mM

Materials

0,0555g CaCl2 (111g/mol)

5mL Sterile Water

Methods

1. Dissolve 0,056g of CaCl2 in 5mL of sterile water
2. Conserve at 4°C

MgCl2 100mM

Materials

0,610g MgCl2 (203,31g/mol)

30mL Sterile Water

Methods

1. Dissolve 0,6105g of MgCl2 in 30mL of sterile water
2. Conserve at 4°C

Sucrose 100mM

Materials

0,171g Sucrose

5mL Sterile Water

Methods

1. Dissolve 0,171g of sucrose in 5mL of sterile water
2. Conserve at 4°C

IPTG 4mM

Materials

80µL IPTG (500mM)

10mL LB

10µL Cam (25ug/mL)

Methods

1. Add 10µL of Cam in 10mL of LB
2. Homogenize
3. Remove 90µL of solution and add 80µL of IPTG
4. Conserve at -20°C

IPTG 4mM

Materials

80µL IPTG (500mM)

10mL LB

10µL Cam (25ug/mL)

Methods

1. Add 10µL of Cam in 10mL of LB
2. Homogenize
3. Remove 90µL of solution and add 80µL of IPTG
4. Conserve at -20°C

Experimentations