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用途
sufficient for ≤40 reactions
包装
pkg of 100 U
制造商/商品名称
Roche
参数
72 °C optimum reaction temp.
储存温度
−20°C (−15°C to −25°C)
一般描述
Pwo SuperYield DNA Polymerase combines the recombinant enzyme Pwo DNA Polymerase with an optimized buffer system. This buffer system enhances the enzymatic properties of the polymerase, resulting in higher yields of the amplification reaction without changing the fidelity of DNA synthesis. This enzyme delivers excellent results due to its enzyme design and optimized buffer system. It amplifies fragments up to 3 kb - even longer amplicons are possible from simple templates.
Pwo DNA Polymerase exhibits increased thermal stability with a half life of greater than 2 hours at +100 °C compared to Taq DNA Polymerase with a half life of less than 5 min at this temperature.
Pwo SuperYield DNA Polymerase, originally isolated from Pyrococcus woesei, is a processive 5′ → 3′ DNA polymerase with 3′ → 5′ exonuclease proofreading activity; 5′ → 3′ exonuclease activity is not detectable.
The enzyme accepts modified nucleotides for efficient labeling of nucleic acids by PCR.
PCR products are blunt-ended directly useable for blunt-end ligation.
Using the magnesium-containing reaction buffer supplied, the final MgCl2 concentration is 1.5 mM.
Pwo DNA Polymerase exhibits increased thermal stability with a half life of greater than 2 hours at +100 °C compared to Taq DNA Polymerase with a half life of less than 5 min at this temperature.
Pwo SuperYield DNA Polymerase, originally isolated from Pyrococcus woesei, is a processive 5′ → 3′ DNA polymerase with 3′ → 5′ exonuclease proofreading activity; 5′ → 3′ exonuclease activity is not detectable.
The enzyme accepts modified nucleotides for efficient labeling of nucleic acids by PCR.
PCR products are blunt-ended directly useable for blunt-end ligation.
Using the magnesium-containing reaction buffer supplied, the final MgCl2 concentration is 1.5 mM.
特异性
Star activity: In cases where star activity is observed and/or the activity of the enzyme in the PCR mix is low, purify the amplification product prior to the restriction enzyme digest using High Pure PCR Product Purification Kit.
应用
Pwo SuperYield DNA Polymerase combines the recombinant enzyme Pwo DNA Polymerase with a newly optimized buffer system. Pwo SuperYield DNA Polymerase is used for the amplification of DNA with the intent to sequence the amplification product or to clone the product (e.g., for the expression of the gene product). The high fidelity of this enzyme makes it particularly suitable for:
- High fidelity PCR
- Site-directed mutagenesis
- Cloning
- Gene expression
- Study of allelic polymorphism in individual RNA transcripts
- Characterization of rare mutations in tissue
- Characterization of the allelic stage of single cells or single DNA molecules
特点和优势
Pwo SuperYield DNA Polymerase yields more high fidelity PCR product. The optimized buffer delivers superior results. Amplify fragments up to 3 kb. A GC-RICH Solution is included for difficult templates. Each dNTPack contains the 10 mM additive-free sodium salt nucleotides in a ready-to-use mix.
- Higher yield and 18-fold higher fidelity
- High performance with difficult templates.
- Reduce working steps in cloning.
- Cost-effective.
包装
1 kit containing 4 components
质量
Each lot is assayed using activated DNA. PCR testing used λ and human genomic DNA.
单位定义
One unit Pwo SuperYield DNA Polymerase is defined as the amount of enzyme that catalyzes the incorporation of 10 nmol total deoxynucleoside triphosphates into acid precipitable DNA within 30 minutes at +70 °C.
Unit Assay: Incubation buffer for assay on activated DNA
20 mM Tris-HCl, pH 8.8 (20 °C), 50 mM KCl, 2.5 mM MgCl2, 10 mM 2-mercaptoethanol, 0.2 mM of each dATP, dCTP, dGTP, dTTP.
Incubation procedure
12.5 mg activated calf thymus DNA and 0.1 mCi [α-32P]dCTP are incubated with 0.01 to 0.1 U Pwo SuperYield DNA Polymerase in 50 μl incubation buffer with a paraffin oil overlay at +70 °C for 30 minutes. The amount of incorporated dNTPs is determined by trichloroacetic acid precipitation followed by scintillation counting.
Volume Activity: 5 U/μl
Unit Assay: Incubation buffer for assay on activated DNA
20 mM Tris-HCl, pH 8.8 (20 °C), 50 mM KCl, 2.5 mM MgCl2, 10 mM 2-mercaptoethanol, 0.2 mM of each dATP, dCTP, dGTP, dTTP.
Incubation procedure
12.5 mg activated calf thymus DNA and 0.1 mCi [α-32P]dCTP are incubated with 0.01 to 0.1 U Pwo SuperYield DNA Polymerase in 50 μl incubation buffer with a paraffin oil overlay at +70 °C for 30 minutes. The amount of incorporated dNTPs is determined by trichloroacetic acid precipitation followed by scintillation counting.
Volume Activity: 5 U/μl
其他说明
仅用于生命科学研究。不可用于诊断。
法律信息
Use of this product is covered by US patent claims and corresponding patent claims outside the US. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patent claims require a separate license from Roche. 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.
仅试剂盒组分
产品编号
说明
- Pwo SuperYield DNA Polymerase 5 U/μl
- PCR Buffer, containing 15 mM MgSO4 10x concentrated
- GC-RICH Solution 5x concentrated
- PCR Nucleotide Mix
危险声明
预防措施声明
危险分类
Aquatic Chronic 3
储存分类代码
12 - Non Combustible Liquids
WGK
WGK 2
闪点(°F)
does not flash
闪点(°C)
does not flash
法规信息
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