LB Agar
LB-Ampicillin Agar
SOB Medium
SOC Medium
TE Buffer
50x TAE
SDS-PAGE
Tris-Glycine-SDS
Protein extraction
A (Wash) buffer
B (Elution) buffer
Protocols:
LB Agar (per Liter)
- ● 10 g of NaCl
- ● 10 g of tryptone
- ● 5 g of yeast extract
- ● 20 g of agar (only for solid media)
- ● Add deionized H2O to a final volume of 1 liter
- ● Adjust pH to 7.0 with 5 N NaOH
- ● Autoclave
LB-Ampicillin Agar (per Liter)
- ● 1 liter of LB agar, autoclaved
- ● Cool to 55°C
- ● Add 10 ml of 10 mg/ml filter-sterilized ampicillin
- ● Pour into petri dishes (~25 ml/100-mm plate)
SOB Medium (per Liter)
- ● 20.0 g of tryptone
- ● 5.0 g of yeast extract
- ● 0.5 g of NaCl
- ● Add deionized H2O to a final volume of 1 liter
- ● Autoclave
- ● Add 10 ml of filter-sterilized 1 M MgCl2 and 10 ml of filter-sterilized 1 M MgSO4 prior to use
SOC Medium (per 100 ml)
- ● Note This medium should be prepared immediately before use.
- ● 2 ml of filter-sterilized 20% (w/v) glucose or 1 ml of filter-sterilized 2 M glucose
- ● SOB medium (autoclaved) to a final volume of 100 ml
TE Buffer
- ● 10 mM Tris-HCl (pH 7.5)
- ● 1 mM EDTA
50 x TAE buffer recipe
- ● 2 M Tris base
- ● Cool to 55°C
- ● 0.05 M of 0.5 M EDTA0
- ● Add deionized H2O water to a final volume of 1 liter
SDS-PAGE Sample 5X Loading Buffer
- ● 250 mM Tris·HCl, pH 6.8
- ● 10% SDS
- ● 30% (v/v) Glycerol
- ● 10 mM DTT
- ● 0.05% (w/v) Bromophenol Blue
- ● Add deionized H2O water to a final volume of 50 ml
Tris-Glycine-SDS running Buffer X10
- ● Dissolve 30.0 g of Tris base
- ● 144.0 g of glycine
- ● 10.0 g of SDS in 1000 ml of H2O
- ● The pH of the buffer should be 8.3
- ● No pH adjustment is required
- ● Store the running buffer at room temperature
- ● Dilute to 1X before use.
Protein extraction lysis buffer
- ● 50 mM Tris·HCl, pH 8.0
- ● 300 mM NaCl
- ● 10 mM Imidazole
- ● 2% Triton X-100
- ● 5% (v/v) Glycerol
- ● 5 mM DTT
- ● 1 mM β-mercaptoethanol
- ● Add deionized H2O water to a final volume of 100 ml
A (Wash) buffer
- ● 50 mM Tris·HCl, pH 8.0
- ● 300 mM NaCl
- ● 10 mM Imidazole
B (Elution) buffer
- ● 50 mM Tris·HCl, pH 8.0
- ● 300 mM NaCl
- ● 500 mM Imidazole