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2GRNTKB

Roche

KAPA2G Robust PCR Kit

Name: KAPA2G Robust PCR Kit
Brand: ROCHE

with dNTPs

別名:

PCR

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About This Item

UNSPSCコード:

41106300

NACRES:

NA.55

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品質水準

100

使用法

sufficient for 200 reactions
sufficient for 500 reactions

シェルフライフ

≤12 mo.

特徴

Difficult Templates/Specialty Enzymes PCR
dNTPs included
hotstart: no

包装

pkg of 100 U (KK5004)
pkg of 250 U (KK5005)

メーカー／製品名

Roche

テクニック

PCR: suitable

入力

crude DNA

保管温度

−20°C

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詳細

The second-generation KAPA2G Robust DNA Polymerase was evolved to solve inconsistent amplification across a broad range of amplicon types (GC- and AT-rich). This product enables higher processivity and specific activity, which translates to robust PCR performance, high sensitivity, and improved tolerance to common inhibitors. The high performance of the KAPA2G Robust DNA Polymerase is ideally suited for challenging PCR applications and difficult samples, eliminating the need for optimization using multiple enzymes and protocols.

アプリケーション

KAPA2G Robust PCR Kit is suitable for:

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

生物化学的／生理学的作用

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.

特徴および利点

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.

品質

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.

調製ノート

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.