Team:UFMG Brazil/Protocols
Project
Lab Work
Modeling
Practices
Synenergene
Team
Protocols
Bacteria mediums
*LB (Luria-Bertani) liquid medium – 300 ml 3 g tryptone 1.5 g yeast extract 3 g NaCl Complete to 300 ml with distilled water Autoclave in a 500 ml erlenmeyer Prepare 2 erlenmeyers with 300 ml of medium each
*LB (Luria-Bertani) liquid medium – 300 ml 3 g tryptone 1.5 g yeast extract 3 g NaCl 4.5 g Agar (weigh directly into the erlenmeyer) Complete to 300 ml with distilled water Autoclave in a 500 ml erlenmeyer *SOC medium – 100 ml 2g tryptone 0.5 g yeast extract 200 µl NaCl 5 M 1 ml MgCl2 1 M 250 µl KCl 1 M 10 ml MgSO4 1 M Complete to 100 ml with distilled water Autoclave and store in bottle with lid
- NaCl MW: 58.44 g/mol
- 58.44 g ---- 1 mol ---- 1000 ml x ------------------------- 50 ml x = 2.92 g
- 2.92 g -------- 1 mol y --------------- 5 mol y = 14.6 g
- MgCl2 MW: 95.211 g/mol
- 95.21 g ---- 1 mol ---- 1000 ml x ------------------------- 50 ml x = 4.7 g
Put 4.7 g MgCl2 into a 50 ml falcon tube and complete to 50 ml with distilled water.
- KCl MW: 74.5513 g/mol
- 74.55 g ---- 1 mol ---- 1000 ml x ------------------------- 50 ml x = 3.72 g Put 3.72 g KCl into a 50 ml falcon tube and complete to 50 ml with distilled water
- Erlenmeyer flask and sterilized becker
- LB Agar
- Sterile petri dishes
- Antibiotic
- Bunsen burner
- 70% Alcohol
- Platinum strap
- Clones
- Ampicillin - 10 µl / ml
- Chloramphenicol -1.2 µL / mL
Bacteria planting
Methods
Heat LB Agar in the microwave. Let it cool near the flame.
Pour between 15mL and 30mL of LB Agar in an Erlenmeyer. Add the antibiotic in the right proportion and mix
Pour the medium in the Petri dish and let it solidify near the flame.
Pipet the desired volume of clone on the plate. Spread the bacterial suspension throughout the petri dish homogeneously.
Cover the plate and incubate it at 37 °C for 12-16
Standard antibiotics concentration used:
