Team:Grenoble-Alpes/Protocols

Lab

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

    CLOSE

    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

    CLOSE

    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

    CLOSE

    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

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    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

    CLOSE

    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

    CLOSE

    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

    CLOSE

    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

    CLOSE

    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

    CLOSE

    Resuspension Solution for Lyophilized Bacteria

    Materials
  • 350µL DMSO
  • 1,5µL 2-mercaptoethanol 1%
  • 5mL Sterile Water

    • Methods
    1. Add 350µL of DMSO and 1,5µL of 2-mercaptoethanol in 5mL of sterile water
    2. Conserve at 4°C

    CLOSE

    Experimentations

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