ALLin™ HS Red Taq Mastermix
highQu ALLin™ Hot Start Taq DNA Polymerase is the superior sensitive enzyme. The activity at room temperature is blocked by small molecular inhibitor.
Detailní popis
Applications
- Hot-start PCR up to 6 kb with a direct gel loading option
- Crude sample PCR
- Low copy target detection
- Amplification of complex (GC/AT rich) templates
- Fast PCR
- TA cloning
- Multiplex hot-start PCR
Benefits
- Outperforming sensitivity & specificity - low copy number target detection and no background
- Higher yields under standard and fast cycling
- Increased sensitivity and success in amplification of longer templates (6 kb), robust amplification of GC rich templates
- Premixed with the red dye and density reagents for direct loading on the gels after the PCR
highQu ALLin™ Hot Start Taq DNA Polymerase is the superior sensitive enzyme. The activity at room temperature is blocked by small molecular inhibitor. Enzyme becomes active only after heating what allows for highly specific and extremely sensitive amplification, no primer dimer formation and no background.
In combination with the optimized ALLin™ buffer enzyme provides higher success rates in demanding PCR applications like amplification of crude, 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.
The convenience of ALLin™ Hot Start Taq DNA Polymerase is maximized by the use of 2X Red colored Mastermix providing the additional advantage of reduced pipetting and minimized errors. The mastermix is even supplied with PCR Water, and the only thing to add is the template with primers.
ALLin™ HS Red Taq Mastermix, 2X is premixed with red dye and density reagents for direct loading on the gels after the PCR.
In a 2% agarose TAE gel the dye migrates with~350 bp DNA, in 1% agarose TAE gel with ~ 600 bp DNA fragments.
ALLin™ HS Red Taq Mastermix, 2X is also a key component in highQu SampleIN™ Direct PCR Kit (DPK0101/5), ensuring outstanding PCR results with crude samples.
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.
- ALLin™ HS Red Taq Mastermix, 2X is premixed with red dye and density reagents for direct loading on the gels after the PCR.
- In a 2% agarose TAE gel the dye migrates with~350 bp DNA, in 1% agarose TAE gel with ~ 600 bp DNA fragments.
Prepare a 50 µl PCR reaction
| Rev. & For. Primers | variable, 0.1-0.4 µM final each (≤ 2 µl of 10 µM) |
| cDNA Template or gDNA Template |
<100 ng or 5 - 500 ng |
| PCR Water | to 25 μl |
| ALLin™HS Red Taq Mastermix, 2X | 25 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) |
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Load probes on the agarose gel. The red loading dye is included in the mastermix.
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After the PCR store probes on ice shortly, for long term storage, keep at -20°C.
| CAT. | SIZE | COMPONENTS | COMPOSITION |
| HSM0301 | 200 r of 50 µl |
5 x 1 ml - ALLin™ HS Red Taq Mastermix, 2X 5 x 1 ml - PCR Water |
1X mastermix contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers, red electrophoresis tracking dye and density reagents for gel loading |
| HSM0305 | 1000 r of 50 µl |
25 x 1 ml - ALLin™ HS Red Taq Mastermix, 2X 25 x 1 ml - PCR Water |
1X mastermix contains 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers, red electrophoresis tracking dye and density reagents for gel loading |
|
Storage |
In the dark at -20°C. |
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