Taq PCR Core Kit

For standard and specialized PCR applications, includes dNTP mix

QIAGEN PCR Buffer for minimal optimization
Additional ready-to-load PCR buffer for faster handling
Q-Solution for amplification of GC-rich templates
Choice of formats for convenience and ease of handling

Taq PCR Core Kit is provided in a complete kit format comprising, Taq DNA Polymerase, the unique QIAGEN PCR Buffer that minimizes the requirement for optimization, as well as a dNTP mix. Also provided is Q-Solution, a novel additive that enables efficient amplification of "difficult" (e.g., GC rich) templates. In addition, CoralLoad PCR Buffer (containing two gel-tracking dyes) is included, enabling immediate loading of PCR products.

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製品名		Cat. no.	List price:
Taq PCR Core Kit (250 U) 250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl₂, dNTP Mix		201223	¥25,500
Taq PCR Core Kit (1000 U) 4 x 250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl₂, dNTP Mix		201225	¥88,500

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Taq PCR Core Kit は分子生物学実験用です。疾病の診断、治療または予防の目的には使用することはできません。

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[A] CoralLoad Concentrate contains two gel-tracking dyes, enabling immediate gel loading of PCR products for [B] easy visualization of DNA migration.|PCR amplification at the indicated Mg²⁺ concentrations using QIAGEN PCR Buffer and Taq DNA Polymerase (QIAGEN). The same PCR was performed in parallel using a PCR buffer and Taq polymerase from another supplier (Supplier A_II). The single-copy human prion protein gene was amplified successfully in each case using the buffer and enzyme from QIAGEN. M: markers.|Two different primer-template systems were amplified in duplicate using QIAGEN PCR Buffer and Taq DNA Polymerase in the absence (–) or presence (+) of 1x Q-Solution. Q-Solution enables specific amplification of difficult templates. [A] human angiotensin receptor II gene; [B] mouse protein kinase C gene; M: markers.|The human single-copy cystic fibrosis gene was amplified usingTaq DNA Polymerase and QIAGEN PCR Buffer using the indicated annealing temperatures. Primers employed were a 22mer with a T_m of 57.5°C (GC content: 54.5%) and a 32mer with a T_m of 85.2°C (GC content: 78%). For analysis, 10% of a 100 µl reaction was loaded. M: markers.|A fragment of the single-copy gene for cystic fibrosis was amplified from 30 ng, 3 ng, and 300 pg human genomic DNA corresponding to 10⁴, 10³, and 10² copies of target template, respectively. Three different lots of Taq DNA Polymerase were used and equal volumes of the PCR product were analyzed on a 1% agarose gel. M: markers.|Three different-sized products from human genomic DNA were amplified using either Taq DNA Polymerase and the QIAGEN PCR Buffer (QIAGEN), or Taq polymerase and a buffer from another supplier (Supplier A_II). For analysis, 10% of each reaction was loaded on the gel. Results from duplicate PCR amplifications are shown. M: markers.|Ammonium and potassium cations in QIAGEN PCR Buffers increase specificity of primer annealing. K⁺binds to the phosphate groups (P^–) on the DNA backbone, stabilizing the annealing of the primers to the template. NH₄⁺, which exists both as the ammonium ion and as ammonia under thermal-cycling conditions, can interact with the hydrogen bonds between the bases (B), destabilizing weak hydrogen bonds at mismatched bases. The combined effect of the two cations maintains the high ratio of specific-to- nonspecific primer-template binding over a wide temperature range.|

Performance

The Taq PCR Core Kit outperformed kits tested from other suppliers and delivers robust PCR performance in a wide range of PCR applications — without the need for time-consuming optimization. The kit includes Taq DNA Polymerase, a high-quality recombinant enzyme that is suitable for general and specialized PCR applications (see figures "Tolerance of different primer T_m Values" and "Specific amplification of long PCR products"). Every lot of Taq DNA Polymerase is subjected to a comprehensive range of quality control tests, including a stringent PCR specificity and reproducibility assay in which low-copy targets are amplified from human genomic DNA (see figure "Lot-to-lot reproducibility"). The unique formulation of QIAGEN PCR Buffer and CoralLoad PCR Buffer, also provided with the kit, enable highly specific PCR in a variety of PCR conditions with minimal optimization requirements (see figures "Wide annealing-temperature window" and "Tolerance to variable magnesium concentration"). In addition, CoralLoad PCR Buffer enables immediate loading of PCR products onto an agarose gel for even easier handling and faster results. Suboptimal PCR can be improved using Q-Solution, a PCR additive, also provided with the kit (see figure "Amplification of difficult templates").

Taq DNA Polymerase specifications

Concentration: 5 units/µl
Recombinant enzyme: Yes
Substrate analogs: dNTP, ddNTP, dUTP, biotin-11-dUTP, DIG-11-dUTP, fluorescent-dNTP/ddNTP
Extension rate: 2–4 kb/min at 72°C
Half-life: 10 min at 97°C; 60 min at 94°C
Amplification efficiency: ≥10⁵ fold
5'–>3' exonuclease activity: Yes
Extra A addition: Yes
3'–>5' exonuclease activity: No
Contaminating nucleases: No
Contaminating RNases: No
Contaminating proteases: No
Self-priming activity: No

Principle

The Taq PCR Core Kit includes everything required for convenient and reliable PCR —Taq DNA Polymerase, QIAGEN PCR Buffer, CoralLoad PCR Buffer, Q-Solution, dNTP Mix, and MgCl₂.

Taq DNA Polymerase

Taq DNA Polymerase is a high-quality recombinant enzyme that is suitable for general and specialized PCR applications (see figures "Tolerance of different primer T_m values" and "Specific amplification of long PCR products").

QIAGEN PCR Buffer

The innovative QIAGEN PCR Buffer has been developed to save time and effort by reducing the need for PCR optimization. QIAGEN PCR Buffer contains both KCl and (NH₄)₂SO₄ (see figure "Increased specificity of primer annealing"). This unique buffer facilitates the amplification of specific PCR products. During the annealing step of every PCR cycle, the buffer allows a high ratio of specific-to-nonspecific primer binding. Owing to a uniquely balanced combination of KCl and (NH₄)₂SO₄, the PCR buffer provides stringent primer-annealing conditions over a wider range of annealing temperatures and Mg²⁺ concentrations than conventional PCR buffers. Optimization of PCR by varying the annealing temperature or the Mg²⁺concentration is dramatically reduced and often not required (see figures "Wide annealing temperature window" and "Tolerance to variable magnesium concentration").

CoralLoad PCR Buffer

CoralLoad PCR Buffer has all the advantages of QIAGEN PCR Buffer. In addition, it can also be used to directly load the PCR reaction onto an agarose gel — separate addition of a gel loading buffer is not required. CoralLoad PCR Buffer provides the same high PCR specificity and minimal reaction optimization as the conventional QIAGEN PCR Buffer. Additionally, it contains two marker dyes — an orange dye and a red dye — that facilitate estimation of DNA migration distance and optimization of agarose gel run time (see figure "CoralLoad PCR Buffer"). The buffer ensures improved pipetting visibility and enables direct loading of PCR products onto a gel, for enhanced convenience.

Q-Solution

Q-Solution facilitates amplification of GC-rich templates or templates with a high degree of secondary structure by modifying the melting behavior of DNA. Use of this unique reagent often enables or improves suboptimal PCR (see figure "Amplification of difficult templates"). Unlike DMSO and other PCR additives, Q-Solution is used at a defined working concentration with any primer–template system and is not toxic.

Procedure

The Taq PCR Core Kit provides all the components required to set up a PCR and can be used for a multitude of PCR-based applications. The optimized, easy-to-follow, streamlined protocol provided with the kit ensures successful PCR results. Suboptimal PCR is simplified with Q-Solution, a unique PCR additive, also included with the kit.

Applications

Taq DNA Polymerase is used for standard and specialized applications, including:

General PCR
RT-PCR
Screening
PCR-based DNA fingerprinting (VNTR, STR, and RAPD)

Feature	Specifications
Applications	PCR, RT-PCR, DNA fingerprinting
dNTP's included	Yes
Enzyme activity	5' -> 3' exonuclease activity
Mastermix	No
Reaction type	PCR amplification
Real-time or endpoint	Endpoint
Sample/target type	Genomic DNA and cDNA
Single or multiplex	Single
With/without hotstart	Without hotstart

パンフレット

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	Analyzing Genetic Differences - (EN) Second edition — innovative tools	詳細を表示
	(EN) - Maximizing PCR and RT-PCR success — Third Edition Addressing critical factors and new solutions	詳細を表示
	(JA) - PCR およびRT-PCR の成功率を高めるために PCR 実験における重要項目と新技術	詳細を表示
	HotStarTaq DNA Polymerase	詳細を表示

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	HotStarTaq DNA Polymerase	詳細を表示

FAQ

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	What should the starting template DNA quality and quantity be for PCR? FAQ ID -74	表示
	What kind of PCR products can be cloned with the QIAGEN PCR Cloning Kit? FAQ ID -165	表示
	How comparable is CoralLoad gel loading dye contained in various QIAGEN PCR Kits to Sigma Red? FAQ ID -1644	表示
	Does QIAGEN sell Q-Solution separately? FAQ ID -204	表示
	How can one determine the optimal annealing temperature for a specific PCR assay? FAQ ID -288	表示
	Is Q-Solution required for PCR with QIAGEN's PCR kits? FAQ ID -380	表示
	Can Taq DNA Polymerase use RNA as a template, and generate false positives in "no-RT" controls? FAQ ID -523	表示
	What is the composition of the QIAGEN 10x PCR Buffer in Taq- and HotStarTaq DNA Polymerase Kits? FAQ ID -606	表示
	Can QIAGEN's Taq- and HotstarTaq DNA Polymerases be used for cycle sequencing? FAQ ID -741	表示
	How much DNA is obtained in the average PCR reaction? FAQ ID -750	表示
	Have you tested the effect of inhibitors on PCR performance? FAQ ID -818	表示
	Do you have a protocol for polyacrylamide gel analysis of oligonucleotides? FAQ ID -961	表示

キットハンドブック

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	Taq PCR Handbook - (EN) For standard and specialized PCR applications with minimal optimization	詳細を表示

テクニカルインフォメーション

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	(EN) - Maximizing end-point PCR success with QIAGEN's automatable PCR solutions	詳細を表示

クイックスタートプロトコール

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	Taq DNA Polymerase and Taq PCR Core Kit (EN)	詳細を表示

MSDS

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	Safety Data Sheets Download Safety Data Sheets for QIAGEN product components.	表示

安全データシート

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	MSDS Taq PCR Core Kit (250 U)
	MSDS Taq PCR Core Kit (1000 U)
	CofA

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CoralLoad PCR Buffer

CoralLoad PCR Buffer ([A]) は、は2種類のゲルマーカー色素を含んでおり ([B ]) 、PCR産物を即ゲルにアプライでき、DNAの泳動を簡単に可視化できる。

異なったマグネシウム濃度への適応

記載のMg²⁺濃度で、QIAGEN PCR BufferとTaq DNA Polymerase（QIAGEN）を用いてPCR反応を行なった。同じ条件のPCR反応を他社（Supplier A_II）のPCRバッファーとTaq DNA ポリメラーゼを用いて行なった。シングルコピーのヒトプリオンタンパク遺伝子が増幅された。M：マーカー。

Amplification of Difficult Templates with Q-Solution

増幅困難なテンプレートを増幅

2種類のプライマー／テンプレートシステムにおいて1x Q-Solutionの非存在下(-)、あるいは存在下(+)でQIAGEN PCR BufferとTaq DNA Polymeraseを用いて増幅した。Q-Solutionを使用すれば、難しいテンプレートを特異的に増幅することができる。[A] ヒトアンジオテンシンレセプターII遺伝子、[B] マウスプロテインキナーゼC遺伝子、M：マーカー。

Tolerance of Different Primer Tm Values.

Tm 値の異なるプライマーへの適応性

ヒト嚢胞性線維症のシングルコピー遺伝子を図に示したアニーリング温度でQIAGEN TaqDNA PolymeraseとPCR Bufferを用いて増幅した。使用したプライマーは、T_m が57.5℃ （GC 含有率：54.5％）のとき22 mer、T_m が85.2℃ （GC 含有率：78％）のとき32 merだった。反応液100 μl の10％をアガロースゲルにロードした。M：マーカー。

ロット間での再現性

ターゲットテンプレート、それぞれ10⁴、10³、10² コピーに相当するヒトゲノムDNA、30 ng、3 ng、300 pgから嚢胞性繊維症のシングルコピー遺伝子のフラグメントを増幅した。Taq DNA Polymeraseの3つの異なるロットを使用し、それぞれのPCR産物は1 ％アガロースゲルで電気泳動を行なった。M：マーカー。

Specific Amplification of Long PCR Products.

長いPCR産物の特異的増幅

Taq DNA PolymeraseとQIAGEN PCR Buffer（QIAGEN）、または他社（Supplier A_II）の TaqDNA ポリメラーゼとバッファーを用いて、ヒトゲノムDNAから異なる長さの3種類のフラグメントを増幅した。分析のため、各反応液の10％をゲルにロードした。同じPCR増幅の結果を示す。M：マーカー。

NH4+ and K+ cations in QIAGEN PCR buffers increase specific primer annealing

プライマーのアニーリングにおける特異性が増加

QIAGEN PCRバッファー中のアンモニウムイオンとカリウムイオンにより、プライマーのアニーリングにおける特異性が増加。K⁺はDNA バックボーンのリン酸基（P^-）に結合し、テンプレートへのプライマーアニーリングを安定化する。サーマルサイクリング条件下では、アンモニウムイオンおよびアンモニアの両方として存在するNH₄⁺は、塩基（B）間の水素結合に相互作用するため、ミスマッチの塩基間の水素結合を不安定にする。これら2 種類の陽イオンの組み合わせ効果により、幅広い温度範囲において非特異的なプライマーテンプレート結合に対する特異的な結合の比率が高く保たれる。