Preparation Potassium Chloride-based PCR Buffer
This guide describes the preparation of Potassium Chloride-based PCR Buffer at a defined molarity for laboratory use.
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Potassium Chloride-based PCR Buffer is commonly used to optimize DNA polymerase activity.
Potassium Chloride-based PCR Buffer – Composition
| Name | Formula | Concentration | CAS |
|---|---|---|---|
| Tris-Cl (pH 9.0) | C₄H₁₁NO₃ | 10 mM | 77-86-1 |
| Potassium chloride | KCl | 50 mM | 7447-40-7 |
| Magnesium chloride | MgCl₂ | 1.5 mM | 7791-18-6 |
| Triton X-100 | C₃₄H₆₂O₁₁ | 0.1% (w/v) | 9002-93-1 |
Sterilization & Storage
- Sterilization: 0.22 μm filtration.
- Storage: Aliquot and store at −20 °C.
Tips
- Buffer pH is usually 8.0–9.5, adjusted by Tris-Cl.
- Triton X-100 is very viscous; use low-concentration stock or substitute with Tween 20.
- When using KCl-based PCR buffer, MgCl₂ is usually at a lower concentration (~1.5 ± 0.25 mM).
- Optimal PCR buffer depends on the DNA polymerase; follow enzyme supplier recommendations.
- Directly using anhydrous MgCl₂ generates heat; add in small portions to partial solvent volume.
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