- Производитель:
- Roche
NACRES: | NA.55 |
Кат. номер |
KK5005 |
KK5004 |
- Amplification of GC- or AT-rich templates
- Templates containing common PCR inhibitors e.g. salts, urea, SDS, or ethanol
- Crude sample PCR e.g. buccal swabs, cultured mammalian, yeast, or bacterial cells
- Colony PCR
- Polymerase chain reaction
Biochem/physiol Actions
KAPA2G Robust DNA polymerase has 5→3 polymerase and 5→3 exonuclease activities, but no 3→5 exonuclease (proofreading) activity. The fidelity of KAPA2G Robust is similar to that of wild-type Taq; it has an error rate of approximately 1 error per 1.7 x 105 nucleotides incorporated. KAPA2G Robust is a highly processive DNA polymerase, capable of amplifying DNA fragments of up to 1 kb in size with a 15 sec/cycle extension time. The use of excessive extension times is likely to result in smearing and nonspecific amplification. For amplicons <1 kb with genomic DNA as a template, 15 sec per cycle should be sufficient for a high yield of the product. In the case of longer amplicons, the time may be increased to a maximum of 60 sec/kb, in 15-sec increments. PCR products generated by KAPA2G Robust are 3-dA-tailed and may be cloned into TA cloning vectors.
Features and Benefits
Efficient amplification of GC- and AT-rich targets
- Robust performance across a wide range of GC- and AT-rich templates
- Increased PCR success rates
Improved tolerance to common PCR inhibitors
- Efficient amplification from crude samples
- Higher yield and sensitivity per unit of enzyme
Unrivalled performance in colony PCR
- Higher yields and improved consistency of PCR direct from E. coli and yeast cells
Quick Notes:
- KAPA2G Robust PCR Kits with dNTPs contain KAPA2G Robust DNA Polymerase, engineered for high processivity and inhibitor tolerance.
- Both purified genomic DNA and crude samples (e.g. colony PCR) can be used as template.
- Use 0.5 U of KAPA2G Robust DNA Polymerase per 25 µL reaction. More challenging PCRs (GCrich,crude sample) may require higher enzyme concentrations.
- Use 15 sec/kb extension time per cycle, and increase to 30-60 sec/kb for difficult amplicons or templates.
- KAPA2G Buffers contain 1.5 mM MgCl2 at 1X.
- Additional MgCl2 may be added using the 25 mM MgCl2 solution provided in the kit.
- KAPA2G Buffer A is optimized for high yield, specificity, and sensitivity.
- KAPA2G Buffer B is recommended for inhibitor contaminated template samples.
- KAPA2G GC Buffer is recommended for GC-rich PCR (>70% GC).
- KAPA Enhancer 1 can be used with KAPA2G Buffer A or B, particularly for GC-rich assays.
- Reaction products are 3-dA-tailed and may be cloned into TA cloning vectors.
Quality
Each batch of KAPA2G Robust DNA Polymerase is confirmed to contain <2% contaminating protein (Agilent Protein 230 Assay). KAPA2G Robust PCR Kits are subjected to stringent quality control tests, are free of contaminating exo- and endonuclease activity, and meet strict requirements with respect to DNA contamination levels.
Preparation Note
Always ensure that the product has been fully thawed and mixed before use. Reagents may be stored at 4°C for short-term use (up to 1 month). Return to -20°C for long-term storage. Provided that all components have been handled carefully and not contaminated, the kit is not expected to be compromised if left (unintentionally) at room temperature for a short period of time (up to 3 days). Long-term storage at room temperature and 4°C is not recommended. Please note that reagents stored at temperatures above -20°C are more prone to degradation when contaminated during use, and therefore storage at such temperatures is at the user′s own risk.
Quality Level | 100, |
shelf life | ≤12 mo. |
feature | Difficult Templates/Specialty Enzymes PCR, dNTPs included, hotstart: no |
packaging | pkg of 100 U (KK5004), pkg of 250 U (KK5005) |
Торговая марка | Roche |
application(s) | PCR: suitable |
input | crude DNA |
shipped in | dry ice |
storage temp. | −20°C |
pictograms | GHS07,GHS08 |
signalword | Warning |
hcodes | H302 - H371 - H412 |
pcodes | P260 - P264 - P270 - P273 - P308 + P311 - P501 |
RIDADR | NONH for all modes of transport |
WGK Germany | WGK 1 |
Flash Point F | does not flash |
Flash Point C | does not flash |
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