ALLin™ Hot Start Taq Polymerase
highQu ALLin™ Hot Start Taq Polymerase is the superior sensitive hot-start DNA polymerase. The enzyme activity at room temperature is blocked by a small molecular inhibitor. Enzyme becomes active only after heating what allows for highly specific and extremely sensitive amplification, no primer dimer formation occurs and no background appears.
Detailní popis
Applications
- Hot-start PCR up to 6 kb, Low copy target detection
- PCR of complex (GC rich) templates
- Fast PCR, multiplexing, TA cloning
Benefits
- Higher yields, no background in standard or fast PCR
- Success on longer (6 kb) or GC rich templates, in crude sample PCR
- 5X ALLin™ PCR Buffer contains optimal Mg2+ and dNTPs
highQu ALLin™ Hot Start Taq Polymerase is the superior sensitive hot-start DNA polymerase. The enzyme activity at room temperature is blocked by a small molecular inhibitor. Enzyme becomes active only after heating what allows for highly specific and extremely sensitive amplification, no primer dimer formation occurs and no background appears.
In combination with the optimized ALLin™ buffer enzyme provides higher success rates in demanding PCR applications like amplification of complex or longer templates and fast cycling. ALLin™ Hot Start Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, 4.5 x 104 (nucleotides incorporated before the error occurs) and produces A-tailed products suitable for ligating into TA cloning vectors.
For the maximum convenience the ALLin™ Hot Start 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™ Hot Start Taq DNA Polymerase, 5 u/µl | 0.25 - 1 µl |
- Mix gently, avoid bubbles.
- Place into the instrument set like:
| Initial denaturation | 1 cycle: 95°C – 1-2 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 shortly on ice, for long term storage keep at -20°C.
| Cat. | Size | Components | Composition |
| HSE0101 | 500 u | 2x 250 u - ALLin™ Hot Start Taq Polymerase, 5 u/µl 4 x 1 ml 5X - ALLin™ PCR Buffer |
Enzyme in storage buffer. 1X ALLin™ PCR Buffer contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers. |
| HSE0105 | 2500 u | 10 x 250 u - ALLin™ Hot Start Taq Polymerase, 5 u/µl 20 x 1 ml - 5X ALLin™ PCR Buffer |
Enzyme in storage buffer. 1X ALLin™ PCR Buffer contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers. |
Storage |
In the dark at -20°C. |
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