How to Prepare 6X DNA Loading Dye
6X concentrated gel loading dye for agarose gel electrophoresis
Components
| Chemical | Concentration |
|---|---|
| Glycerol | 30% v/v |
| Bromophenol blue | 0.25% w/v |
| Xylene cyanol FF | 0.25% w/v |
| EDTA | 10 mM (pH 8) |
Preparation Strategy
Method: stock_then_dilute
Reason: EDTA mass (0.29g) < 1g threshold; bromophenol blue and xylene cyanol masses are very small (<0.3g each). Stock approach ensures accuracy and avoids weighing errors.
Preparation Table
| Component | Conc. | Amount | Unit |
|---|---|---|---|
| EDTA disodium salt dihydrate | 10 mM final | 1.0 mL | mL |
| Glycerol | 30% v/v | 30.0 | mL |
| Bromophenol blue | 0.25% w/v | 250 | mg |
| Xylene cyanol FF | 0.25% w/v | 250 | mg |
| Water | Balance | to 100 | mL |
Procedure
- 1Step 1: Prepare 1M EDTA stock (pH 8.0): Dissolve 2.92g EDTA disodium salt dihydrate in ~8mL water, adjust pH to 8.0 with NaOH pellets, bring to 10mL final volume
- 2Step 2: In a 100mL graduated cylinder, add 60mL nuclease-free water
- 3Step 3: Add 250mg bromophenol blue powder slowly while stirring vigorously to ensure complete dissolution
- 4Step 4: Add 250mg xylene cyanol FF powder and mix thoroughly until completely dissolved
- 5Step 5: Add 30.0mL glycerol and mix well
- 6Step 6: Add 1.0mL of 1M EDTA stock (pH 8.0) and mix
- 7Step 7: Adjust final volume to 100mL with nuclease-free water
- 8Step 8: Mix thoroughly by inversion until homogeneous
- 9Step 9: Check that solution is dark blue-purple color with no precipitate
pH Adjustment
pH should be ~8.0 from EDTA stock - do not require further adjustment. EDTA provides buffering capacity.
Sterilization
Filter through 0.22 μm syringe filter into sterile tubes. Do not autoclave as dyes may degrade and EDTA may precipitate.
Safety
Level 1 safety: Standard lab coat, safety glasses, nitrile gloves. Dyes can stain - avoid skin contact. Work in well-ventilated area when handling powders.
Storage
Store at 4°C in dark bottles or wrap in foil to protect from light. Aliquot into small volumes (1-5mL) to minimize freeze-thaw cycles. Stable for 1 year. Do not freeze as glycerol may crystallize.
Need a different volume or concentration?
Customize this solutionRelated Solutions
TE Buffer
Standard DNA/RNA storage buffer
20X SSC Buffer
Standard hybridization and wash buffer for Southern/Northern blots
Church Hybridization Buffer
Rapid hybridization buffer for Southern/Northern blots
DEPC-Treated Water
RNase-free water for RNA work, autoclave after DEPC treatment to decompose residual DEPC
10X MOPS Running Buffer
10X stock for denaturing RNA agarose gel electrophoresis
RNA Loading Buffer
Denaturing loading buffer for RNA agarose gels