ALLin™ TaqDNA Polymerase
highQu ALLin™ Taq DNA Polymerase is the versatile engineered enzyme which in combination with the optimized ALLin™ buffer provides higher success rates in demanding PCR applications like amplification of complex templates, crude sample PCR and fast cycling.
Detailní popis
Applications
- Routine PCR up to 6 kb
- PCR of GC rich templates
- Colony PCR, Fast PCR, TA cloning
Benefits
- Higher yields under standard and fast cycling
- Increased success in amplification of longer (6 kb) and GC rich templates
- 5X ALLin™ PCR Buffer contains optimal Mg2+ and dNTP concentrations
highQu ALLin™ Taq DNA Polymerase is the versatile engineered enzyme which in combination with the optimized ALLin™ buffer provides higher success rates in demanding PCR applications like amplification of complex templates, crude sample PCR and fast cycling. ALLin™ Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, 4.5 x 104 (a number of correct nucleotides incorporated before the first error). ALLin™ Taq DNA Polymerase produces A-tailed products suitable for ligating into TA cloning vectors. For the maximum convenience the ALLin™ Taq Mastermix, 2X is available.
Important Notes
- Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips.
- Include a no-template control and positive control in parallel.
- Thaw and keep reagents on ice. Mix well before use.
- The longer the amplicon, the longer the extension time: 15 sec/kb - <6kb and 1 sec/kb - <1kb. Use >90 sec extension for multiplexing
- Run an annealing temperature gradient from 55°C to 65°C to choose the best specificity conditions.
- Do not use fast cycling for multiplexing.
Prepare a 50 µl PCR reaction
| Rev. & For. Primers | variable, up to 0.4 µM final each (2 µl of 10 µM each) |
| cDNA Template or gDNA Template |
<100 ng or 5 - 500 ng |
| 5X ALLin™ PCR Buffer | 10 μl |
| Water | to 49 μl |
| ALLin™ Taq DNA Polymerase, 5u/μl | 0.25 - 1 µl |
- Mix gently, avoid bubbles.
- Place into the instrument set like:
| Initial denaturation | 1 cycle: 95°C - 1 min |
| Denaturation | 40 cycles: 95°C - 15 sec |
| Annealing | 40 cycles: 55-65°C – 15 sec |
| Extension | 40 cycles: 72°C – 1- 90 sec (15 sec/kb) |
After the PCR store probes on ice shortly, store at -20°C for long term.
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