Rotor-Gene SYBR^® Green PCR Kit

SYBR Green を用いたRotor-Geneサイクラーでの超高速リアルタイムPCRおよび2ステップRT-PCR

低コピーターゲットでも特異的で高感度な検出
幅広いテンプレート量で正確に検出
至適化なしに信頼できる結果を実現する特別調製のマスターミックス
QuantiTect Primer Assayと組み合わせて最適な結果

Rotor-Gene SYBR Green PCR KitはRotor-Gene QおよびRotor-Geneサイクラーでの使用を目的としてデザインされ、SYBR Green I検出を用いたリアルタイム定量PCRあるいは2ステップリアルタイムRT-PCR で特異性の高いｇDNAやcDNAターゲットの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 SYBR Green PCR Kit (400) For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene SYBR Green PCR Master Mix, 2 x 2 ml RNase-Free Water		204074	411,00 €	Add to cart

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The Rotor-Gene SYBR^® Green 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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Highly specific and sensitive detection.|Reliable and specific detection down to 10 copies.|Specific primer annealing.|Fast primer annealing.|

Real-time two-step RT-PCR was carried out using tenfold dilutions of human leukocyte cDNA (10 ng to 0.1 ng) and a QuantiTect Primer Assay for BCL2 (B-cell CLL/lymphoma 2). Reactions were run in triplicate using either [A] the Rotor-Gene SYBR^® Green PCR Kit and Rotor-Gene Q or [B] a kit and cycler from Supplier A_II. The Rotor-Gene system provided lower C_T values and greater reproducibility within triplicates. Melting curve analysis (see insets) showed a single peak, demonstrating specific amplification of the target.|Real-time PCR was carried out using tenfold dilutions of plasmid DNA (10⁹ to 10 copies) and a QuantiTect Primer Assay for PPIA (cyclophilin A). Reactions were run in triplicate using the Rotor-Gene SYBR^® Green PCR Kit and Rotor-Gene Q. [A] 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. [B] Melting curve analysis indicates specific amplification of the target.|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 SYBR^® Green 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 SYBR Green PCR KitはcDNAターゲットの遺伝子解析で使用するのに最適です（図“特異性と感度の高い検出” および“10コピーのターゲットを確実かつ高い特異性で検出”)。

QuantiTect Primer AssaysはRotor-Gene SYBR Green PCR Kit と組み合わせると、至適化なしに特異的なPCR 産物の高感度な定量が行なえます（表参照）。

SYBR Green を用いたRT-PCR で卓越した性能
	QIAGEN		A_II社
	C_T	平均偏差	C_T	平均偏差
BAX (BCL2結合Xタンパク質)	24.84	0.05	29.57	0.46
BCL2 (アポトーシス遺伝子)	26.96	0.05	32.83	0.29
MYC (がん原遺伝子)	28.42	0.21	35.26	0.72
β-アクチン（ハウスキーピング遺伝子）	20.24	0.03	24.39	0.12

Principle

Rotor-Gene SYBR Green PCR Kit は Rotor-Gene Q において反応条件やサイクリング条件の至適化なしに信頼できる迅速なリアルタイムPCR定量を実現します。マスターミックス中の蛍光色素SYBR Green Iは、ターゲットに特異的な標識プローブを合成しなくても多数の異なるターゲットを解析できます。特異的なプライマーアニーリングを促進するK⁺とNH₄⁺イオンのバランスの取れた配合により、特異性の高い増幅が確保され、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 SYBR Green 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ｑPCRのセットアップ
Rotor-Gene SYBR Green 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 SYBR Green PCR Buffer	ユニークな添加物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
SYBR Green I 色素	ニ本鎖DNAに結合して強い蛍光シグナルを発生	感度の高い定量

Procedure

Rotor-Gene SYBR Green 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テンプレートとプライマー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。

２ステップリアルタイムRT-PCRを用いた遺伝子発現解析において、 Rotor-Gene SYBR Green PCR Kit、 QuantiTect Reverse Transcription Kit、QuantiTect Primer Assays、Rotor-Gene Q の組み合わせは、最適化なしに開始できるトータルソリューションです。QuantiTect Reverse Transcription Kitは、ゲノムDNAコンタミの除去を組み合わせたcDNA合成をわずか20分で行ないます。QuantiTect Primer Assay は、ヒト、マウス、ラット、その他の生物種のほとんどの遺伝子に対応するバイオインフォマティックスで検証済みのプライマーセットです。アッセイはGeneGlobeウェブポータルサイトで容易に注文できます。

Applications

Rotor-Gene SYBR Green PCR KitはRotor-Gene Q上でcDNAやgDNAターゲットの迅速なリアルタイム定量に最適です。本キットはまたRotor-Gene 3000およびRotor-Gene 6000でも使用可能です。

Rotor-Geneサイクラ―上でSYBR Green Iを用いてRNAターゲットを超迅速な1ステップでqRT-PCRで遺伝子解析する場合には、Rotor-Gene SYBR Green 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 SYBR Green I on Rotor-Gene cyclers
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	SYBR Green I
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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	What is a QuantiTect Primer Assay? FAQ ID -1141	View
	How do you achieve fast cycling, yet still deliver the same performance in PCR as that achieved with standard cycling? FAQ ID -1430	View
	Do you have a protocol for Rotor-Gene software setup for the Rotor-Gene SYBR Green PCR and RT-PCR Kits? FAQ ID -2104	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
	Why is a 2-step (and not a 3-step) cycling protocol recommended for Rotor-Gene SYBR Green Kits? FAQ ID -2122	View
	Do QuantiTect Primer Assays also work with Rotor-Gene SYBR Green PCR Kits? FAQ ID -2123	View
	Will QuantiTect Primer Assays work with Rotor-Gene SYBR Green Kits using an annealing step at 60ºC? FAQ ID -2124	View
	Do you have any information or guidelines regarding the choice of reference genes for real-time PCR? FAQ ID -2371	View
	How do I create a workspace that is free of DNA contamination, prior to carrying out a qPCR experiment? FAQ ID -2654	View
	What real-time cycler should I use for my qPCR experiments? FAQ ID -2670	View
	What is a dissociation curve, and why is it important to run a dissociation curve, following qPCR using SYBR Green chemistry? FAQ ID -2678	View
	What is the threshold cycle or Ct value? FAQ ID -2682	View
	How can I predict the percent qPCR signal due to contaminating DNA, for a given qPCR assay, and its matching NRT control? FAQ ID -2688	View
	Why do I see multiple high-intensity peaks in my qPCR dissociation curve at temperatures less than 70 ºC? FAQ ID -2690	View

Kit Handbooks

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	(EN) - Rotor-Gene SYBR Green Handbook For fast real-time PCR, two-step RT-PCR, and one-step RT-PCR using SYBR Green I on Rotor-Gene cyclers	Show details
	(EN) - QuantiTect Primer Assay Handbook For genomewide, ready-to-use real-time RT-PCR assays using SYBR Green detection	Show details

Technical Information

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

Supplementary Protocols

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	Rotor-Gene® software setup for the Rotor-Gene SYBR® Green PCR Kit - (EN)	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 SYBR Green PCR Kit (400)

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Highly specific and sensitive detection.

Reliable and specific detection down to 10 copies.

Real-time PCR was carried out using tenfold dilutions of plasmid DNA (10⁹ to 10 copies) and a QuantiTect Primer Assay for PPIA (cyclophilin A). Reactions were run in triplicate using the Rotor-Gene SYBR^® Green PCR Kit and Rotor-Gene Q. [A] 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. [B] Melting curve analysis indicates specific amplification of the target.

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 SYBR^® Green 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.

Rotor-Gene SYBR® Green PCR Kit

Rotor-Gene SYBR^® Green PCR Kit