Rotor-Gene Probe PCR Kit

配列特異的プローブを用いた超高速リアルタイム定量PCRおよび2ステップRT-PCR用

低コピー数の標的遺伝子でも高感度な検出
幅広いテンプレート量で正確に検出
Rotor-Gene サイクラーでの信頼できる結果を超高速で実現するために最適化済み
高速サイクリング用に特別に調製された即使用可能なマスターミックス

Rotor-Gene Probe PCR Kitは、Rotor-Gene Qおよびその他のRotor-Gene サイクラーでの使用を目的としてデザインされ、配列特異的プローブを用いたリアルタイムPCRや2ステップリアルタイムRT-PCRでgDNAおよびcDNAの超高速で高感度な定量を実現します。特別に至適化されたマスターミックスとRotor-Geneサイクラーの遠心エアコントロール方式の組み合わせにより、最高の性能を実現します。マスターミックスは2～8 ℃で保存でき、簡便に取り扱えます。

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Product		Cat. no.	List price:
Rotor-Gene Probe PCR Kit (400) For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene Probe PCR Master Mix, 2 x 2 ml RNase-Free Water		204374	$920.00	Add to cart

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The Rotor-Gene Probe PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

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Real-time PCR was carried out using two-fold dilutions of human genomic DNA (30 ng to 3.75 ng) and a self-designed TaqMan assay for IL1R2 (interleukin 1 receptor, type II). Reactions were run using either [A] the Rotor-Gene Probe PCR Kit and Rotor-Gene Q or [B] a kit and cycler from Supplier A_II (5 replicates per template dilution). The Rotor-Gene system provided lower C_T values, greater reproducibility within replicates, and greater resolution of different template dilutions.|Real-time PCR analysis of human genomic DNA was carried out using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. [A] Analysis of two-fold dilutions of DNA (from 30 ng [5000 copies] to 0.06 ng [20 copies]) using a self-designed TaqMan assay for IL1R2 (interleukin 1 receptor, type II); 5 replicates per template dilution. The average difference in C_T value between adjacent template dilutions was 1.07 cycles. [B] Analysis of 5 ng DNA using a self-designed TaqMan assay for IL1R2. From 100 replicates, the standard deviation was 0.21. [C] Analysis of 20 ng DNA using a self-designed TaqMan assay for ACTB (actin, beta). From 100 replicates, the standard deviation was 0.29.|Real-time PCR was carried out using fourfold dilutions of human genomic DNA (equivalent to 16, 666 cells down to 1 cell) and a self-designed TaqMan assay for ACTB (actin, beta). Reactions were run in triplicate using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. The Rotor-Gene system provided reliable detection over the entire range of template dilutions, with high reproducibility within each set of triplicates.|Real-time PCR was carried out using ten-fold dilutions of plasmid DNA (10⁹ to 10 copies) and a self-designed TaqMan assay for PPIA (cyclophilin A). Reactions were run in triplicate using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. The Rotor-Gene system provided reliable detection over the entire range of template dilutions as well as highly reproducible C_T values within each set of triplicates.|Cations in the Rotor-Gene Q PCR buffer increase specific primer annealing. K⁺ binds to phosphate groups on double-stranded DNA, stabilizing primer annealing. NH₄⁺ destabilizes weak hydrogen bonds between mismatched bases.|[A] Q-Bond in Rotor-Gene Probe PCR Master Mix increases the affinity of DNA polymerase for short single-stranded DNA, reducing primer annealing time to a few seconds. In addition, the unique buffer composition supports the melting of DNA, reducing denaturation and extension times. [B] Without Q-Bond, the primer and polymerase bind sequentially to the template, increasing primer annealing time.|

Performance

Rotor-Gene Probe PCR Kitは定量PCR解析に最適で（図“高感度で再現性のある検出”、“精度の高いPCR”)、幅広いテンプレート量で信頼できる検出を実現します（表および図 “精度の高いPCR”、“1個の細胞まで確実に検出”、“幅広いダイナミックレンジ”)。

幅広いダイナミックレンジにわたり信頼できる検出
コピー	平均C_T	C_T 差
1,000,000,000	5.11	3.07
100,000,000	8.18	3.39
10,000,000	11.57	3.04
1,000,000	14.61	3.68
100,000	18.29	3.28
10,000	21.57	3.68
1,000	25.25	3.06
100	28.31	3.43
10	31.74	–
効率	0.99
直線性	1

Principle

Rotor-Gene Probe PCR KitはRotor-Gene Qにおいて信頼できる2ステップリアルタイム定量RT-PCRを実現します。反応条件やサイクリング条件の至適化は不要です。本キットは加水分解プローブ（例、TaqManプローブ）を使用するようにデザインされています。特異的なプライマーアニーリングを促進するK⁺/NH₄⁺イオンのバランスの取れた組み合わせが特異性の高い増幅を確保し、PCRの高い特異性と感度を実現します (図 “プライマーの特異的なアニーリング”)。画期的なPCR 添加物であるQ-Bond により性能を損なうことなく高速サイクリングが行なえ、ラン時間はわずか45 分です（図“プライマーの高速なアニーリング”）。

2x Rotor-Gene Probe PCR Kitの成分*
成分	特長	利点
HotStarTaq Plus DNA Polymerase	95℃、5分の活性化	室温での定量PCRのセットアップ
Rotor-Gene Probe PCR Buffer	NH₄⁺とK⁺イオンの配合バランス	特異性の高いプライマーのアニーリングで信頼性の高いｑPCR結果
Rotor-Gene Probe PCR Buffer	ユニークな添加剤Q-Bond	より迅速なPCRランタイムでより早く結果が得られ、一日あたりの反応数も増大

Procedure

即使用可能なRotor-Gene Probe PCR Master Mixにより、反応条件やサイクリング条件の至適化が不要です。DNAテンプレート、プライマー、プローブをマスターミックスに添加し、サイクラーをプログラミングします。キットに添付のハンドブックに詳細な説明が記載されています。

加水分解プローブ（例、TaqManプローブ）は、Rotor-Gene Probe PCR Kitと一緒にRotor-Gene Qで使用でき、マスターミックスにプライマー/プローブミックスとテンプレートを添加するだけで迅速で高感度な定量を実現します。

2ステップリアルタイムRT-PCRで良好な結果を得るためには、QuantiTect Reverse Transcription Kitを用いてcDNAを合成することをお薦めします。ゲノムDNAの除去を組み合わせて、わずか20分で迅速なcDNA合成を実現します。

Applications

Rotor-Gene Probe PCR Kitは、Rotor-Gene Q上で配列特異的なプローブを用いたgDNA/cDNAターゲットの迅速なリアルタイムPCRや2ステップリアルタイムRT-PCR用キットです。本キットはRotor-Gene 3000およびRotor-Gene 6000サイクラーにも対応しています。

Rotor-Geneサイクラー上で配列特異的なプローブを用いてRNAターゲットを超高速な1ステップqRT-PCRで遺伝子発現解析する場合には、Rotor-Gene Probe RT-PCR Kitをご使用ください。

Feature	Specifications
Applications	Real-time quantification of genomic DNA or cDNA targets
Description	For ultrafast quantitative real-time PCR and two-step RT-PCR using sequence-specific probes
Reaction type	Real-time PCR and two-step RT-PCR
Real-time or endpoint	Real-time
Sample/target type	DNA, cDNA
Single or multiplex	Single
SYBR Green I or sequence-specific probes	Sequence-specific probes
Thermal cycler	Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000
With or without ROX	Without ROX dye

Brochures & Guides

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	(EN) - Rotor-Gene Q - Pure Detection Now with even more applications!	Show details

FAQs

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	How do you achieve fast cycling, yet still deliver the same performance in PCR as that achieved with standard cycling? FAQ ID -1430	View
	Are Rotor-Gene Kits compatible with reaction setup using the QIAgility instrument? FAQ ID -2116	View
	Do the master mixes in Rotor-Gene Kits contain dUTP to allow UNG pretreatment? FAQ ID -2117	View
	Why is the initial activation step different for Rotor-Gene Probe, SYBR Green and Multiplex Kits? FAQ ID -2118	View
	What are the main differences between Rotor-Gene and QuantiTect or QuantiFast PCR Kits? FAQ ID -2119	View
	Will Rotor-Gene Kits also work on the Rotor-Gene 6000 and 3000 cyclers? FAQ ID -2121	View
	How do Rotor-Gene Probe Kits compare with QuantiFast Probe Kits? FAQ ID -2125	View
	Are TaqMan® Gene Expression Assays from Applied Biosystems compatible with Rotor-Gene Probe Kits? FAQ ID -2126	View
	Do you have any information or guidelines regarding the choice of reference genes for real-time PCR? FAQ ID -2371	View
	What real-time cycler should I use for my qPCR experiments? FAQ ID -2670	View
	What is the threshold cycle or Ct value? FAQ ID -2682	View

Kit Handbooks

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	(EN) - Rotor-gene Probe Handbook For fast real-time PCR, two-step RT-PCR, and one-step RT-PCR using sequence-specific probes on Rotor-gene cyclers.	Show details

Technical Information

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	(EN) - Critical factors for synthesis of cDNA for real-time PCR analysis	Show details

Safety Data Sheets

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

Safety Data Sheets

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	MSDS Rotor-Gene Probe PCR Kit (400)

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Highly sensitive and reproducible detection.

High precision in real-time PCR.

Real-time PCR analysis of human genomic DNA was carried out using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. [A] Analysis of two-fold dilutions of DNA (from 30 ng [5000 copies] to 0.06 ng [20 copies]) using a self-designed TaqMan assay for IL1R2 (interleukin 1 receptor, type II); 5 replicates per template dilution. The average difference in C_T value between adjacent template dilutions was 1.07 cycles. [B] Analysis of 5 ng DNA using a self-designed TaqMan assay for IL1R2. From 100 replicates, the standard deviation was 0.21. [C] Analysis of 20 ng DNA using a self-designed TaqMan assay for ACTB (actin, beta). From 100 replicates, the standard deviation was 0.29.

Reliable detection down to 1 cell.

Real-time PCR was carried out using fourfold dilutions of human genomic DNA (equivalent to 16, 666 cells down to 1 cell) and a self-designed TaqMan assay for ACTB (actin, beta). Reactions were run in triplicate using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. The Rotor-Gene system provided reliable detection over the entire range of template dilutions, with high reproducibility within each set of triplicates.

Wide dynamic range.

Real-time PCR was carried out using ten-fold dilutions of plasmid DNA (10⁹ to 10 copies) and a self-designed TaqMan assay for PPIA (cyclophilin A). Reactions were run in triplicate using the Rotor-Gene Probe PCR Kit and Rotor-Gene Q. The Rotor-Gene system provided reliable detection over the entire range of template dilutions as well as highly reproducible C_T values within each set of triplicates.

Specific primer annealing.

Cations in the Rotor-Gene Q PCR buffer increase specific primer annealing. K⁺ binds to phosphate groups on double-stranded DNA, stabilizing primer annealing. NH₄⁺ destabilizes weak hydrogen bonds between mismatched bases.

Fast primer annealing.

[A] Q-Bond in Rotor-Gene Probe PCR Master Mix increases the affinity of DNA polymerase for short single-stranded DNA, reducing primer annealing time to a few seconds. In addition, the unique buffer composition supports the melting of DNA, reducing denaturation and extension times. [B] Without Q-Bond, the primer and polymerase bind sequentially to the template, increasing primer annealing time.