REF: EG25104-M/L

Storage Condition

−20℃

Components

Component	EG25104M	EG25104L
2× Taq PCR Aurora Mix	5 × 1 ml	100 × 1 ml

Description

2× Taq PCR Aurora Mix is a ready-to-use PCR master mix that minimizes hands-on time and contamination risk. Products carry a 3′-A overhang for direct T/A cloning after purification.

The mix contains Taq DNA polymerase and a proofreading protein with 3′→5′ exonuclease activity to robustly amplify targets up to ~7 kb across broad GC contents (≈30–70%). An optimized buffer boosts performance and the extension rate is ~2–4× faster than WT-Taq, shortening total run time.

Built-in blue and red tracking dyes allow direct gel loading without adding separate loading buffer. Dyes do not affect amplification; for optical downstream analyses (absorbance/fluorescence), purify PCR products first.

Quality Control Assays

Endonuclease Activity: 200 ng supercoiled plasmid + 25 μl mix, 4 h @ 37℃ → <10% nicked/linearized.
Non-specific Nuclease Activity: 15 ng dsDNA fragments + 25 μl mix, 16 h @ 37℃ → no detectable degradation.

Protocol

1) PCR Reaction System (50 μl)

Reagent	Amount	Final
2× Taq PCR Aurora Mix*	25 μl	1×
Forward Primer (10 μM)	1–2 μl	0.2–0.4 μM
Reverse Primer (10 μM)	1–2 μl	0.2–0.4 μM
Template DNA	x μl	—
ddH₂O	to 50 μl	—

* Thaw on ice before use. Typical template amounts (per 50 μl): genomic DNA 10–200 ng; plasmid/viral DNA 10 pg–5 ng.

2) Three-Step PCR Program

Step	Temp	Time
Initial Denaturation†	95℃	3–5 min
Denaturation	95℃	30 s
Annealing‡	55–65℃	30 s
Extension§	72℃	15–30 s/kb
Final Extension	72℃	5 min

30–35 cycles.

3) Two-Step PCR Program

Step	Temp	Time
Initial Denaturation†	95℃	3–5 min
Denaturation	95℃	30 s
Annealing + Extension‡§	60–65℃	30 s/kb
Final Extension	72℃	5 min

† For colony PCR, 95℃ for ~10 min helps lyse E. coli/S. cerevisiae.
‡ Set annealing by primer Tm; raise if specificity is poor. Temperature gradient helps optimization.
§ <3 kb: 15 s/kb; >3 kb: ~30 s/kb. For highest yield, use a uniform 30 s/kb.

Notice

Primer Design Tips

Prefer G/C at the 3′ terminal base; avoid 3′ hairpins.
Match primer Tm values within ~1℃; target 55–65℃ (check with software such as Primer Premier).
GC content ~40–60%; avoid long homopolymers and high GC/AT regions.
Avoid ≥5 nt internal complementarity or ≥3 nt 3′ complementarity between primers.
BLAST to confirm specificity.

Electrophoresis & Staining

Post-stain gels (soak method) for clearer indicator bands. In 1% TAE, the blue dye migrates ~1,500 bp; the red dye ~100 bp.

FAQ & Troubleshooting

Problem	Possible Reason	Solution
No/low product	Primer design; annealing temp; primer conc.; short extension; low cycles; template quality/amount	Redesign primers; gradient to find optimal T_a; 0.2–0.4 μM primers; 30 s/kb extension; 35–40 cycles; use cleaner template / adjust input
Non-specific bands/smear	Primer issues; low T_a; high primer conc.; too long extension; too many cycles; template impurities/excess	Redesign; increase T_a toward 65℃ in 2℃ steps; 0.2 μM primers; shorten extension; 25–30 cycles; clean template / adjust input

Write a review

Your Name

Your Review

Note: HTML is not translated!

Rating Bad Good

Captcha

Please complete the captcha validation below

2×Taq PCR Aurora Mix

Brand: Best Enzymes
Product CAT#: EG25104M
Availability: In Stock

Available Options

Enzymes Handling Fee

Overnight Shipping Handling (+$29.99)

Size

Qty

0 reviews / Write a review