Griffitts:Common buffers
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Electrophoresis Buffers
2% bromophenol blue
Sterile filter
8X DNA loading
- 40% glycerol
- 1/50 dilution of bromophenol blue
6X SDS loading
- 7.5 mL 80% glycerol
- 2.5 mL 1M Tris pH 6.8
- 1.2 g SDS
- 600 μL 2-mercaptoethanol
- 150 μL 2% bromophenol blue
DNA ladder
- 147 μL ddH20
- 34 μL 8X DNA loading buffer
- 20 μL 1 kb ladder from NEB
- 2 μL 0.25M EDTA
Note: use 5 μL per lane; 3kb band=60 ng
Protein ladder
- Open vial of Sigma SDS6H2-1VL
- Add 1 mL ddH20
- Add 500 μL 6X SDS loading
- Aliquot 50 μL per tube and store at -20
Note: use 5 μL per lane
ethidium bromide (10 mL)
- 10 mL ddH20
- 10 mg ethidium bromide
Note: use 8 μL per 50 mL gel
50X TAE (1 L)
- ~800 mL ddH20
- 242 g Trizma base
- 57.1 mL glacial acetic acid
- 100 mL 0.25 M EDTA (pH 8.0)
QS to volume with ddH20
Note: this is half the amount of EDTA compared to standard TAE
10X Laemmli running buffer
QS to volume with ddH20
Coomassie stain (300 mL)
- 120 mL ethanol
- 0.3 g Brilliant Blue R
Mix for 30 min.
- 30 mL acetic acid
- 150 mL ddH20
Coomassie destain solution (1 L)
- 550 mL ddH20
- 400 mL ethanol
- 50 mL acetic acid
DNA extraction buffers
colony lysis solution (for PCR) (100 mL)
- 96.2 mL of sterile ddH2O
- 0.5 mL of 1 M Triz pH 8.0 (final concentration: 5 mM)
- 0.8 mL of .25 M EDTA (final concentration: 2 mM)
- 2.5 mL of 20% Triton X-100 (final concentration: 0.5%)
- in fridge with the antibiotics
plasmid purification solutions
resuspension buffer
lysis solution
neutralization solution
- 4 M KOAc pH 5.5 (acetic acid)
RNAse A
silica suspension
Silicon dioxide is purchased from Sigma (S5631)
Wash 5X with ddH20
Resuspend as 50% slurry in ddH20