产品说明

一般描述

PCR involves replication of a DNA template by a thermostable DNA polymerase. The processivity, specificity, and fidelity of the polymerase enzyme used can influence the efficiency, reproducibility, and yield of the PCR reaction. High-fidelity PCR, utilizes a DNA polymerase with a low error rate and results in a high degree of accuracy in the replication of the DNA of interest. Fidelity is critical when accurate sequence amplification of the gene target is needed, for example, when direct sequencing or cloning for downstream protein expression. Unwarranted mutation could severely impact your studies. Our analysis has shown that KOD enzymes are an easy choice for fast, accurate and high-yielding PCR. EMD Millipore′s molecular biologists work to develop and formulate polymerases offering the highest specificity, fidelity and yield during PCR amplification. In addition, optimized buffer compositions, convenient master mixes and cycling parameters provide additional ease of use and data reproducibility.KOD Hot Start DNA Polymerase* is a premixed complex of KOD DNA Polymerase and two monoclonal antibodies that inhibit the DNA polymerase and 3′→5′ exonuclease activities at ambient temperatures (Mizuguchi 1999). KOD Hot Start amplifies genomic DNA templates up to 21 kb including GC-rich genes for PCR applications. KOD Hot Start combines the high fidelity, fast extension speed, and outstanding processivity of KOD with the high specificity of an antibody-mediated hot start. Non-specific amplification is reduced because mispriming events that can occur during setup and initial temperature increase are avoided. In addition, primer degradation during setup at room temperature due to exonuclease activity is effectively inhibited. KOD Hot Start DNA Polymerase generates blunt-ended PCR products suitable for cloning with the Novagen Perfectly Blunt^® and LIC Vector Kits.
Source: Recombinant Thermococcus kodakaraensis KOD1 DNA polymerase expressed in E. coli
Concentration: 1.0 U/l
Nicking activity: None detected
Amplification effiency: Functional PCR; inhibition of activity at 21℃ verified
Storage: -20℃
*Manufactured by Toyobo and distributed by EMD. Not available in Japan.
Note: Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser¢s own internal research. No other patents rights (such as 5¢ Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California, 94404, USA.

应用

KOD Hot Start DNA Polymerase has been used:

• in the amplification of genomic DNA with high fidelity for sanger sequencing.
• for genomic DNA amplification to screen single-cell clones for homozygous deletion of the targeted exon.
• for the amplification of specific gene families on the genomic DNA isolated from mice lymphoma cells.

包装

200 u in Fibre case
1000 u in Glass bottle

特点和优势

• High Fidelity: Approximately 80-fold higher fidelity than conventional Taq DNA polymerase
• Long & Accurate PCR: Amplifies genomic DNA up to 12 kbp and plasmid and lambda DNA templates up to 21 kbp with low error rate
• Efficiently amplifies high GC-rich regions
• Antibody-mediated hot start feature reduces mispriming and primer-dimer formation
• Produces blunt end PCR products suitable for cloning
• Convenient room-temperature setup compatible with automation
• Compatible with site-directed mutagenesis protocols
• Optimal KOD Hot Start Buffer for PCR performance over a wide range of targets

组分

•200 U 或 5 × 200 UKOD Hot Start DNA聚合酶(1.0 U/µl)

•1.2 ml 或 5 × 1.2 ml用于KOD Hot Start DNA聚合酶的10X PCR缓冲液

•1 ml 或 5 × 1 ml25 mM MgSO₄

•1 ml 或 5 × 1 mldNTP 混合物(各2 mM)

警告

毒性：标准处理(A)

单位定义

在含有20 mM Tris-HCl(pH 7.5，25°C)、8 mM氯化镁₂、7.5 mM DTT、50 µg/ml BSA、分别150 µM dATP、dCTP、dGTP、dTTP(未标记形式和[³H] dTTP的混合物)、以及150 µg/ml活化的小牛胸腺DNA的反应中，一个单位定义为在75°C、30分钟内可催化10 nmol dNTP掺入酸不溶形式所需要的酶量。

其他说明

Mizuguchi, H., et al. 1999.J. Biochem.(Tokyo)126, 762.Kitabayashi, M., et al. 2002.Biosci.Biotechnol.Biochem.66 (10), 2194.Fujii, S., et al. 1999.J. Mol.Biol.289, 835.

法律信息

由Toyobo制造并由EMD分销。购买本产品包括豁免根据产品说明书中所规定的专利提起的诉讼，仅可将所购买的量用于购买者自己的内部研究。′其他专利权(例如5′核酸酶工艺的专利权)不得明示、暗示或禁止反言。有关购买许可的更多信息，可联系Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA获取。
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany
Perfectly Blunt is a registered trademark of Merck KGaA, Darmstadt, Germany

产品性质

安全信息

象形图
警示用语：	Warning
危险声明	H319
预防措施声明	P264 - P280 - P305 + P351 + P338 - P337 + P313
危险分类	Eye Irrit. 2
储存分类代码	10 - Combustible liquids
WGK	WGK 2