Made up to 400mL with Milli-Q Water in a 500mL Schott bottle. Autoclaved for 15 minutes to sterilize.
LB Agar (400mL)
Tryptone
4g
Yeast Extract
2g
NaCl
4g
Bacto Agar
6g
Made up to 400mL with Milli-Q water in a 500mL in a Schott bottle. Autoclaved for 15 minutes to sterilize. 400µL of antibiotic was added once the bottle was cool enough to handle. Plates were poured using antiseptic technique.
SOB Medium (400mL)
Tryptone
8g
Yeast Extract
2g
5M NaCl
0.8mL
1M KCl
1mL
1M MgCl2
4mL
1M MgSO4
4mL
Made up to 400mL with distilled water. Autoclaved for 15 minutes to sterilize.
SOC Medium
Tryptone
8g
Yeast Extract
2g
5M NaCl
0.8mL
1M KCl
1mL
1M MgCl2
4mL
1M MgSO4
4mL
2M D-Glucose
4mL
Made up to 400mL with Milli-Q water. Autoclaved for 15 minutes to sterilize.
1% Agarose Gel
DNA Grade Agarose
1g
1x TAE Buffer
100mL
GelRed
4µL
Mixed components in 250mL flask and microwaved until completely dissolved. Cooled slightly before 4µL of GelRed added.
M9 Glycerol Agar
M9 Salts (5x)
100mL
1M MgSO4
2mL
1M CaCl2
50µL
20% Glycerol
10mL
Milli-Q Water
113mL
The next reagents were added to the mixture via filter sterilization:
1% Thiamine HCl
15mL
Cas Amino Acids
=10mL
Plates were then prepared using aseptic technique.
M9 Glucose Agar
Agar solution was prepared by autoclaving 7.5g of agar in 250mL of water.
The following reagents were combined and autoclaved before being aseptically added to the hot agar:
M9 Salts (5x)
100mL
1M MgSO4
2mL
1M CaCl2
50µL
20% Glucose
10mL
Milli-Q Water
113mL
The next reagents were added to the mixture via filter sterilization:
1% Thiamine HCl
15mL
Cas Amino Acids
10mL
Plates were then prepared using aseptic technique.
