Team:Lyon-INSA-ENS/Realisation/Protocols

General culture conditions

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Antibiotics were used at the following concentrations :
Ampicilin (Amp) : 100µg/mL
Chloramphenicol (Cm) : 20µg/mL
Spectinomycin (Spc) : 100µg/mL
Kanamycin (Kan) : 30µg/mL
The solvent is a 70/30 (v/v) water/ethanol mix for chloramphenicol, other antibiotics were dissolved in water. All volumes mentionned are meant for a 100X mother solution.

The composition of the different media we have used is the following :
LB : 10 g of tryptone, 5 g of yeast extract, and 10 g of NaCl for 1L of liquid LB. Solid LB for plates is made by adding agar to reach a concentration of 1.5%.
M63G : 100/1/1 (v/v) of M63, glucose 20% and LB
LB/2 : 50/50 (v/v) mix of LB and water

Choose one of our protocols to read its description

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. Biofilm quantification

. CaCl2 chemical transformation

. Fermentas digestion

. Ligation

. NucleoSpin Plasmid QuickPure : Isolation of high-copy plasmid DNA from E. coli

. Ozyme digestion

. Plasmid or Cosmid DNA purification using HiSpeed Plasmid Midi kits

. Plasmid DNA purification using QIAprep® Spin Miniprep Kit

. Pouring of plates

. Storage of strains

. TSS chemical transformation