EquiPhi29™ DNA Polymerase

Citations & References (18)

Thermo Scientific™

EquiPhi29™ DNA Polymerase

Name: EquiPhi29™ DNA Polymerase
SKU: A39390

Thermo Scientific EquiPhi29 DNA Polymeraseは、in vitroタンパク質の進化によって開発された特許取得済みのphi29 DNAポリメラーゼ変異です。この酵素は、タンパク質の熱安定性詳細を見る

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製品番号（カタログ番号）	数量	ProductName
A39390	250 U、10 U/μL
A39391	1000 U、10 U/μL
A39392	5000 U、10 U/μL

3 オプション

製品番号（カタログ番号） A39390

価格（JPY）

14,600

Each

お問い合わせください ›

数量:

250 U、10 U/μL

一括またはカスタム形式をリクエストする

Thermo Scientific EquiPhi29 DNA Polymeraseは、in vitroタンパク質の進化によって開発された特許取得済みのphi29 DNAポリメラーゼ変異です。この酵素は、タンパク質の熱安定性、反応速度、製品収量、増幅バイアスにおいてphi29 DNAポリメラーゼよりも大幅に改善されており、高い処理能力（70 kb以上）、強力なストランド置換活性、一本鎖DNAまたはRNAに優先的に作用する3'→5'エキソヌクレアーゼ（プルーフリーディング）活性など、野生型酵素のすべての利点を保持しています。そのため、エキソ耐性のあるランダムプライマーが推奨されます。

主な特長：
•市場で提供される最小の増幅バイアス
• 少量のテンプレートからでも非常に高い増幅されたDNA収量
• より短時間での高精度なDNA合成

アプリケーション：
•さまざまなサンプルタイプを使用した全ゲノムのバイアスなし増幅（WGA）
   –単一細胞からのDNA
   –培養されていない微生物細胞とウイルス粒子
   –病原体またはメタゲノム
   –SNPおよびSTRの検出のためのDNA
• ローリングサークル増幅（RCA）
• タンパク質がプライミングされたDNA増幅
• 致死性のDNAの無細胞ローニング
• パドロックプローブによるin situジェノタイピング
• RNAがプライミングされたDNA増幅

注：EquiPhi29 DNAポリメラーゼで反応混合物にピロホスファターゼを添加すると、DNA合成が向上する場合があります。

研究用途にのみご使用ください。診断目的には使用できません。

仕様

濃度10 U/μL

最適反応温度42°C

反応時間2 hours

出荷条件ドライアイス

使用対象（アプリケーション）MDA-WGA, RCA

ポリメラーゼEquiPhi29 DNA ポリメラーゼ

製品タイプStand-alone enzyme

数量250 U、10 U/μL

Unit SizeEach

組成および保存条件

• 25 µL EquiPhi29 DNAポリメラーゼ（10 U/µL）
• 0.25 mLのEquiPhi29 DNAポリメラーゼ反応バッファー（10倍）
• 0.25 mLのDTT（100 mM）

-5～-30℃で保存してください。

よくあるご質問（FAQ）

What is the error rate of EquiPhi29 DNA Polymerase?

The error rate of EquiPhi29 DNA Polymerase is 6 x 10^-6.
The error rate of EquiPhi29 DNA Polymerase was measured according to the method described in literature:
Mielinis, P., Sukackaitė, R., Serapinaitė, A., Samoilovas, F., Alzbutas, G., Matjošaitis, K., & Lubys, A. (2021). MUA-based molecular indexing for rare mutation detection by Next-Generation sequencing. Journal of Molecular Biology, 433(19), 167209. https://doi.org/10.1016/j.jmb.2021.167209

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Can I use EquiPhi29 DNA Polymerase to incorporate 5-methyl-dCTP?

Thermo Scientific EquiPhi29 DNA Polymerase is a proprietary mutant phi29 DNA Polymerase developed through in vitro protein evolution. EquiPhi29 DNA Polymerase as well as phi29 DNA Polymerase should be able to incorporate 5-methyl-dCTP nucleotides and other modified nucleotides.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

When using EquiPhi29 DNA Polymerase, do I need to purify amplified DNA products before downstream applications?

Cleaning of the amplified product is not required prior to several downstream methods (e.g., debranching, digestion with restriction endonucleases, Sanger sequencing); the dilution of amplified product is sufficient. If the clean-up procedure is needed, we recommend using an affinity-based spin-column or magnetic bead-based purification method.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

What is the minimal recommended time for amplification with EquiPhi29 DNA Polymerase?

The optimal reaction time for DNA amplification with EquiPhi29 DNA Polymerase is 2 hours. For samples with ≥1 pg of DNA input, DNA amplification time can be shortened to 1 hour if maximizing product yield is not essential.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Can liquid media culture or colonies be used as a starting material for amplification with EquiPhi29 DNA Polymerase?

Yes. EquiPhi29 DNA Polymerase can work with different types of sample input material such as purified DNA, liquid media culture, agar plate colonies, etc.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

View all EquiPhi29™ DNA Polymerase, 250 U FAQs

引用および参考文献 (18)

引用および参考文献

Abstract

Fidelity of phi 29 DNA polymerase. Comparison between protein-primed initiation and DNA polymerization.

Authors:Esteban JA, Salas M, Blanco L

Journal:J Biol Chem

PubMed ID:8428945

'Phi 29 DNA polymerase is able to catalyze two different synthetic reactions: protein-primed initiation and DNA polymerization. We have studied the fidelity of phi 29 DNA polymerase when carrying out these two reactions. Global fidelity was dissected into three steps: insertion discrimination, mismatch elongation, and proofreading. The insertion discrimination of ... More

Whole-metagenome amplification of a microbial community associated with scleractinian coral by multiple displacement amplification using phi29 polymerase.

Authors:Yokouchi H, Fukuoka Y, Mukoyama D, Calugay R, Takeyama H, Matsunaga T

Journal:Environ Microbiol

PubMed ID:16817924

'Limitations in obtaining sufficient specimens and difficulties in extracting high quality DNA from environmental samples have impeded understanding of the structure of microbial communities. In this study, multiple displacement amplification (MDA) using phi29 polymerase was applied to overcome these hindrances. Optimization of the reaction conditions for amplification of the bacterial ... More

Suitability of genomic DNA synthesized by strand displacement amplification (SDA) for AFLP analysis: genotyping single spores of arbuscular mycorrhizal (AM) fungi.

Authors:Gadkar V, Rillig MC

Journal:J Microbiol Methods

PubMed ID:15936100

'Limited biological samples of microbial origin often yield insufficient amounts of genomic DNA, making application of standard techniques of genetic analysis, like amplified fragment length polymorphism (AFLP), virtually impossible. The Phi29 DNA polymerase based whole genome amplification (WGA) method has the potential to alleviate this technical bottleneck. In the present ... More

Novel application of Phi29 DNA polymerase: RNA detection and analysis in vitro and in situ by target RNA-primed RCA.

Authors:Lagunavicius A, Merkiene E, Kiveryte Z, Savaneviciute A, Zimbaite-Ruskuliene V, Radzvilavicius T, Janulaitis A

Journal:RNA

PubMed ID:19244362

We present a novel Phi29 DNA polymerase application in RCA-based target RNA detection and analysis. The 3'-->5' RNase activity of Phi29 DNA polymerase converts target RNA into a primer and the polymerase uses this newly generated primer for RCA initiation. Therefore, using target RNA-primed RCA, padlock probes may be targeted ... More

Towards the analysis of the genomes of single cells: further characterisation of the multiple displacement amplification.

Authors:Panelli S, Damiani G, Espen L, Micheli G, Sgaramella V

Journal:Gene

PubMed ID:16564650

The development of methods for the analysis and comparison of the nucleic acids contained in single cells is an ambitious and challenging goal that may provide useful insights in many physiopathological processes. We review here some of the published protocols for the amplification of whole genomes (WGA). We focus on ... More

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