Features a 5–15 s/kb amplification rate and produces 3'-A overhangs for direct TA cloning. Ideal for routine PCR and colony PCR
Кот. Нет. :
Y101311-1MLБренд :
YanbiotechПроисхождение продукта :
ChinaОбъем :
1 mLProduct 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 | 1× |
| (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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@ Wuhan Yanbiotech Co., Ltd. Все права защищены
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