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2×Fast sTaq PCR Master Mix

2×Fast sTaq PCR Master Mix

Features a 5–15 s/kb amplification rate and produces 3'-A overhangs for direct TA cloning. Ideal for routine PCR and colony PCR

  • Cat. No. :

    Y101311-1ML
  • Brand :

    Yanbiotech
  • Product Origin :

    China
  • Volume :

    1 mL
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  • Product Details

Product Information:

This ready-to-use PCR premix (2×) contains modified Taq DNA polymerase, dNTPs, and an optimized reaction buffer. Designed for routine simple template PCR and colony/cell PCR. Simply add template, primers, and ddH₂O to a 1× final concentration. Offers strong amplification capacity with fast extension rates of 5–15 s/kb, and produces 3'-A overhangs. Pre-mixed with Loading Dye for direct agarose gel electrophoresis.


Instructions for Use:

Recommended PCR reaction system

 Component  20 μL 50 μL Final Concentration
 Templatea  Variable Variable as required
 Forward Primer (10 μM)b  0.8 μL 2 μL 0.4μM
 Reverse Primer (10 μM)b  0.8 μL 2 μL 0.4 μM
 2×Fast High Fidelity PCR Master Mix  10 μL 25 μL
 (DMSO, optional)c  (0.6 μL) (1.5 μL) (3%)
ddH2O Add to 20 μL Add to 50 μL  
 

a: Template

1. Plasmid or phage DNA: 50 ng–5 pg per 50 μL reaction

2. Genomic DNA: 250 ng–50 ng per 50 μL reaction

3. cDNA: Dilute 2–100 fold; add ≤10% of total reaction volume

4. Bacterial culture or crude sample: Add ≤10% of reaction volume

5. Excess template → non‑specific amplification; insufficient template → low amplification efficiency

b: Primer

1. Final concentration: 0.2–1.0 μM (recommended: 0.4 μM)

2. Too little → low yield or no amplification; excess → non‑specific amplification
c: High-GC Template
3. DMSO: Up to 10% can be added to the reaction mixture

Recommended PCR amplification conditions

Step Temperature Volume Cycles
Initial Denaturationa 98℃ 30 s-120 s 1
Denaturation 98℃ 5-10 s 25-35
Annealingb 50-72℃ 10-30 s
Extensionc 72℃ 5-15 s/kb
Final extension 72℃ 5-10 min 1
Hold 4-16℃ Forever  

a: A pre-denaturation time of 30 s is suitable for most conventional templates; complex templates can be denatured for up to 2 min.

b: For amplification of complex templates, containing concatenated primers, the annealing time can be extended up to 30 s.

c: The recommended extension speed of plasmid and other simple templates is 5-10 s/KB; The recommended extension speed of conventional genomic template is 10-15 s/KB; Complex template 15-30 s/KB.


Notice: 

1. Thoroughly thaw and mix before use. It can be kept at 4℃ for at least 2 weeks to avoid freeze-thaw cycles.

2. For your safty and health, please wear safety glasses, gloves, or protective clothing.

3. For Research Use Only

 

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