How to Prepare 1x TAE Buffer
Standard agarose gel electrophoresis buffer for DNA, pH ~8.5
Components
| Chemical | Concentration |
|---|---|
| Tris base | 40 mM |
| Acetic acid | 20 mM |
| EDTA | 1 mM (pH 8) |
Preparation Strategy
Method: direct_dissolution
Reason: All components can be directly dissolved as masses are >1g or volumes are reasonable
Preparation Table
| Component | Conc. | Amount | Unit |
|---|---|---|---|
| Tris base | 40 mM | 4.85 | g |
| Glacial acetic acid | 20 mM | 1.14 | mL |
| EDTA disodium salt dihydrate | 1 mM | 0.37 | g |
Procedure
- 1Step 1: Add approximately 800 mL of distilled water to a 1L beaker or flask
- 2Step 2: Add 4.85 g Tris base and stir until completely dissolved
- 3Step 3: Slowly add 1.14 mL glacial acetic acid while stirring continuously
- 4Step 4: Add 0.37 g EDTA disodium salt dihydrate
- 5Step 5: If EDTA does not dissolve completely, add NaOH pellets one at a time until dissolved
- 6Step 6: Adjust final volume to exactly 1L with distilled water
- 7Step 7: Mix thoroughly and verify pH is approximately 8.3
pH Adjustment
pH should naturally be ~8.3. If adjustment needed, use NaOH to raise or additional acetic acid to lower. EDTA dissolution may require pH >8.
Sterilization
Autoclave at 121°C, 15 psi for 25 minutes (1L volume). Allow to cool completely before use.
Safety
Level 2 safety: Wear nitrile gloves, lab coat, and safety glasses. Use fume hood when adding glacial acetic acid. EDTA powder should be handled in fume hood to avoid inhalation.
Storage
Store at room temperature in tightly sealed container. Stable for 6 months. No refrigeration required. Check for precipitates before use.
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