- Производитель:
- Roche
NACRES: | NA.55 |
Кат. номер |
KK5702 |
KK5701 |
- amplification of GC- or AT-rich templates
- aplification of templates containing common polymerase chain reaction (PCR) inhibitors e.g. salts, urea, SDS, or ethanol
- crude sample PCR e.g. buccal swabs, cultured mammalian, yeast, or bacterial cells
- colony PCR
- DNA extraction and amplification
- mutation screening
- methylation-specific PCR (MSP) amplification
- genotyping-PCR
- PCR-based sequencing
Biochem/physiol Actions
DNA fragments generated with KAPA2G Robust HotStart® DNA Polymerase have the same characteristics as DNA fragments generated with wild-type Taq DNA polymerase and may be used for routine downstream analyses or applications, including restriction enzyme digestion, cloning, and sequencing. Like wild-type Taq, KAPA2G Robust HotStart has 53 polymerase and 53 exonuclease activities, but no 35 exonuclease (proofreading) activity. The fidelity of KAPA2G Robust HotStart is similar to that of wild-type Taq; it has an error rate of approximately 1 error per 1.7 x 105 nucleotides incorporated. PCR products generated by KAPA2G Robust HotStart 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 HotStart ReadyMix PCR Kits 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 15 sec/kb extension time per cycle, and increase to 30-60 sec/kb for difficult amplicons or templates.
- KAPA2G Robust HotStart ReadyMix contains 2 mM MgCl2 at 1X.
- KAPA2G Robust HotStart ReadyMix with dye includes two inert tracking dyes to allow direct loading onto agarose gels.
- KAPA2G Robust HotStart ReadyMix is ideal for single-protocol PCR, i.e. amplification of fragments varying from 25–65% GC, up to 1 kb in size, using a single reaction setup and cycling protocol.
- Reaction products are 3-dA-tailed and may be cloned into TA cloning vectors.
Quality
Each batch of KAPA2G Robust HotStart DNA Polymerase is confirmed to contain <2% contaminating protein (Agilent Protein 230 Assay). KAPA2G Robust HotStart ReadyMix 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.
ReadyMix is a trademark of Sigma-Aldrich Co. LLC
shelf life | 12 mo. |
Quality Level | 100, |
feature | Difficult Templates/Specialty Enzymes PCR, dNTPs included, hotstart |
packaging | kit of 1.25 mL (100 x 25 µL rxn; KK5701), kit of 6.25 mL (500 x 25 µL rxn; KK5702) |
Торговая марка | Roche |
concentration | 2 x ( contains 2 mM MgCl2 at 1x) |
application(s) | PCR: suitable |
input | crude DNA |
shipped in | dry ice |
storage temp. | 20°C |
pictograms | GHS07,GHS08 |
signalword | Warning |
hcodes | H302 - H371 |
pcodes | P260 - P264 - P270 - P301 + P312 + P330 - 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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