ALLin™ Taq Mastermix
highQu ALLin™ Taq DNA Polymerase je všestranně navržený enzym, který v kombinaci s optimalizovaným pufrem ALLin™ poskytuje vyšší míru úspěšnosti v náročných aplikacích PCR, jako je amplifikace komplexních templátů, PCR surového vzorku a rychlé cyklování.
Detailní popis
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 error occurs) and produces A-tailed products suitable for ligating into TA cloning vectors.
The convenience of ALLin™ Taq DNA Polymerase is maximized by the use of 2X Mastermix providing the additional advantage of reduced pipetting and minimized errors. The mastermix is even supplied with the PCR Water, and the only thing to add is the template with primers.
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 |
| PCR Water | to 25 μl |
| ALLin™ Taq Mastermix, 2X | 25 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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