FlamePol Master Mix Ready to Load MgCl2 (mM)

2. Mix Composition (5x)

• FlamePol DNA polymerase
• 5x Reaction Buffer: 0.4 M Tris-HCl, 0.1 M (NH4​)2​SO4​, 0.1% w/v Tween-20
• 7.5 mM MgCl2​ $\to$ in 1×: 1.5 mM MgCl2​
• 1 mM dNTPs of each $\to$ in 1×: $200\mu \text{M}$ dATP, $200\mu \text{M}$ dCTP, $200\mu \text{M}$ dGTP, $200\mu \text{M}$ dTTP
• Blue dye (migrates like a 3.5–4.5 kb fragment)
• Yellow dye (migrates like 35–45 bp)
• Compound that increases sample density for direct loading.

3. Usage Recommendations

• Recommended for any PCR that will be visualized by agarose gel electrophoresis and ethidium bromide staining.
• Not recommended for applications that rely on spectrophotometric measurements (absorbance or fluorescence), as the blue and yellow dyes may interfere.
• To prevent contamination, set up reactions under a laminar flow hood or in a PCR box.

4. Recommended PCR reaction mix (20 μL total)

5. Recommended PCR cycling protocol

The extension time depends on the length of the fragment to be amplified. A time of 1 min/kb is recommended.

6. Safety

Handle only by trained personnel. Wear a lab coat, gloves, and safety glasses. Avoid contact with skin/eyes; in case of contact, wash with water. Consult the Safety Data Sheet (SDS).

7. Shipping and Storage

• Routine storage: −18 ∘C to −28 ∘C.
• Shipping and temporary storage: up to 1 month at room temperature or up to 6 months at 2–8 ∘C, with no detrimental effects on product quality.
• FOR RESEARCH USE ONLY, NOT FOR USE IN DIAGNOSTIC PROCEDURES.