Storage Condition

−20℃

Components

Description

Taq-Plus PCR Forest Mix (2×) V2 is a pre-mixed PCR reaction solution containing Taq DNA Polymerase, dNTPs, and buffer components—everything required for PCR except DNA template and primers. The premix reduces pipetting steps, saving time and minimizing contamination during setup.

The mix supports robust amplification of targets up to 6 kb from genomic DNA. Products carry a 3′ A-overhang and are directly suitable for T/A cloning.

Forest Mix includes two tracking dyes for direct gel loading. Dyes do not interfere with PCR, but purify products before absorbance/fluorescence measurements. A dye-free Taq-Plus PCR Master Mix (2×) V2 is also available.

Quality Control Assays

Endonuclease Activity

50 μl reaction with 200 ng supercoiled plasmid + 25 μl Forest Mix, 4 h at 37℃ → <10% nicked/linearized (agarose gel).

Non-specific Nuclease Activity

50 μl reaction with 15 ng dsDNA fragments + 25 μl Forest Mix, 16 h at 37℃ → no detectable degradation (agarose gel).

1) PCR Reaction System (50 μl)

^a Recommended 0.2–0.4 μM; adjustable 0.1–1 μM.

Template guidance (per 50 μl): genomic DNA ~10–400 ng; plasmid/viral DNA ~10 pg–20 ng.

2) PCR Conditions (30–35 cycles)

^a For complex templates (e.g., bacterial/yeast colony), extend to ~10 min to improve denaturation.

^b Use 30 s/kb for <3 kb targets; 60 s/kb for >3 kb.

3) Migration Distance of Dyes vs. Gel Concentration

Note: dyes can affect absorbance measurements—purify PCR product if needed.