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Team:Evry/Protocols/04
From 2013.igem.org
Solutions, Media and Petri Dishes
Antibiotics
Carbenicillin:Put 150 mg of Carbenicilline in 15 mL of distilled water (10 mg per mL).
Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -4 °C.
Use 1 µL of Carbenicillin for 1 mL of solution.
Kanamycin:
Put 750 mg of Kanamycin in 15 mL of distilled water (50 mg per mL).
Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -4 °C.
Use 1 µL of Kanamycin for 1 mL of solution.
Chloramphenicol:
Put 525 mg of Kanamycin in 15 mL of ethanol (35 mg per mL).
Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -4 °C.
Use 25 µL of Chloramphenicol for 1 mL of solution.
Carbenicillin inhibit cell wall synthesis, kanamycin interfer with mRNA traduction and chloramphenicol inhibit ribosomial peptidyl transferase activity.
LB and LB Agar
Mix 20 g of LB Broth in 1L of distilled water in a bottle of 1L, then put in autoclave.Mix 17,5 g of LB Agar and in 500 mL of distilled water in a bottle of 1L, then put in autoclave.
CaCl2 and CaCl2+glycerol
Solution for 1M Cacl2:Add 14,30g of CaCl2 into 100 ml desalted water
Solution for 0,1M Cacl2:
Add 50 mL of CaCl2 1M solution into 450 ml of desalted water
Solution for 0,1M Cacl2 + 15% glycerol:
Add 50 mL of CaCl2 1M solution and 75 mL of glycerol 100% into 450 ml of desalted water
Tris HCl
Disolve 4,6g of Trisbase in 30 mL of distilled water.Adjust pH with concentrated HCl(~4 mL) until pH=7,5.
Add distilled water until 50 mL then put in autoclave.
If the solution has a yellow coloration, do the preparation again with better Trisbase.
Tris HCl is used to store DNA.
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