QuantiFast Multiplex RT-PCR Kits
配列特異的なプローブを用いた遺伝子発現解析用高速1ステップマルチプレックスリアルタイム定量RT-PCR
- 同一ウェル内で複数のターゲットRNAを高感度で検出
- 時間を最大50%まで短縮し迅速な結果
- 至適化実験なしに良好なマルチプレックスRT-PCRを実現
- わずかなターゲット量の違いも正確に判別
- リファレンス遺伝子と3種類までのターゲットを同一チューブ内で検出可能
QuantiFast Multiplex RT-PCR Kit は、1ステップのマルチプレックスリアルタイム定量RT-PCR により、同一チューブ中で最高4種類のRNAターゲットを高速かつ確実に定量します。特殊な割合で配合された逆転写酵素により迅速で効率的なcDNA合成を実現します。Q-bondテクノロジーと至適化済みのマスターミックスにより、短いRamping Time機能を持つ高速サイクラーだけではなくほとんどのサイクラーで性能を損なうことなく高速なマルチプレックスリアルタイムRT-PCRを行なえます。ホットスタートと即使用可能なマスターミックス中の独自のPCRバッファーシステムの組み合わせは、最適化実験の必要なしにほとんどのサイクラーで高感度な定量RT-PCRを実現します。2種類のキットが入手できます:蛍光補正のためにROXが必要なサイクラー用QuantiFast Multiplex RT-PCR Kitおよびその他のサイクラー用のQuantiTect Multiplex RT-PCR +R Kit。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。
en-US
Configure
Added to your shopping cart
|
Produto
|
Product no.
|
Num Cat.
|
Preço de lista:
|
|
|
Este produto foi descontinuado.
Em vez disso, recomendamos
|
|
Show details
|
|
|
varies
|
select
Can't order online?
To place an order via phone, email or for requesting a quote, please provide the product’s name, number and catalog number.
|
|
|
| Produto |
Num Cat. |
Preço de lista: |
|
|
Produto
|
Num Cat.
|
Preço de lista:
|
|
|
QuantiFast Multiplex RT-PCR Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast Multiplex RT-PCR Master Mix (with ROX dye), 100 µl QuantiFast RT Mix, 2 x 2 ml RNase-Free Water
|
204854
|
|
|
|
QuantiFast Multiplex RT-PCR +R Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast Multiplex RT-PCR Master Mix (without ROX dye), 100 µl QuantiFast RT Mix, 210 µl ROX Dye Solution, 2 x 2 ml RNase-Free Water
|
204954
|
|
|
|
QuantiFast Multiplex RT-PCR +R Kit (2000)
For 2000 x 25 µl reactions: 25 ml 2x QuantiFast Multiplex RT-PCR Master Mix (without ROX dye), 500 µl QuantiFast RT Mix, 1.05 ml ROX Dye Solution, 1 x 20 ml RNase-Free Water
|
204956
|
|
|
QuantiFast Multiplex RT-PCR Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Significantly reduced RT-PCR times.|Reliable relative quantification.|Comparable results in triplex and singleplex RT-PCR.|Uncompromised sensitivity in 4-plex RT-PCR.|Fast primer annealing.|QIAGEN multiplex kits.|Unique buffer promotes stable and efficient primer annealing.|
QuantiFast Multiplex Kits reduce total RT-PCR run time by up to 50% in real-time one-step RT-PCR (40 cycles run; comparison with QuantiTect Multiplex Kits). I: iCycler iQ; L1: LightCycler 480; L2: LightCycler 2.0; A1: ABI PRISM 7900; A2: Applied Biosystems 7500 Fast System; M: Mx3005P. |Cell line X was treated with one of two compounds (Y or Z) or left untreated (U). RNA was purified and duplex, real-time one-step RT-PCR was carried out on the Applied Biosystems 7500 Fast System using the QuantiFast Multiplex RT-PCR +R Kit and TaqMan Gene Expression Assays for myogenin and GAPDH. For each treatment, 5 independent experiments were performed. [A] Changes in myogenin expression were detected with high accuracy and reproducibility (green curves). The expression of the housekeeping gene GAPDH was similar in all experiments (blue curves), allowing normalization of myogenin expression levels using the ΔΔCT method of relative quantification. [B] The fold changes in normalized myogenin expression level relative to that in untreated cells were consistent between experiments, as indicated by the small error bars. (Data kindly provided by Angelika Meyer, Novartis Pharma AG, Basel, Switzerland).|Triplex and singleplex, real-time one-step RT-PCR were carried out on the LightCycler 480 using the QuantiFast Multiplex RT-PCR +R Kit and self-designed TaqMan assays for [A] RPS27A (a ribosomal protein), [B] GAPDH (a housekeeping gene), and [C] UBC (a housekeeping gene). The template was Ramos cell line RNA (10 ng, 1 ng, or 0.1 ng), and reactions were run in duplicate. The comparable CT values for triplex PCR (colored curves) and singleplex PCR (gray curves) and [D] the high PCR efficiencies demonstrate the reliability of triplex PCR with the QuantiFast Multiplex RT-PCR +R Kit when analyzing targets of differing abundance.|4-plex, real-time one-step RT-PCR was carried out on the Mx3005P using the QuantiFast Multiplex RT-PCR +R Kit and Primer Express designed TaqMan assays. Duplicate reactions were run using 10 ng RNA from Ramos cells as template. All 4 targets, which varied greatly in abundance, were reproducibly detected.|[A] Q-Bond in QuantiFast Buffer allows the DNA polymerase and primer to bind as a single complex, reducing the 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 annealing time.|QuantiFast Multiplex RT-PCR Kits provide a simple procedure for quantitative, multiplex, real-time PCR. In contrast to current methods, the kits eliminate the need for optimization of the concentrations of primers, Mg2+, and Taq DNA polymerase, even for difficult assays (e.g., assays in which the copy number of the target gene is much smaller than that for the reference gene).|[A] NH4+ ions prevent nonspecific primers from annealing to the template. [B] Synthetic Factor MP, an innovative PCR additive, increases the local concentration of primers at the template. Together with K+ and other cations, Factor MP stabilizes specifically bound primers, allowing efficient primer extension by HotStarTaq Plus DNA Polymerase.|
Desempenho
QuantiFast Multiplex RT-PCR KitはRT-PCR時間を最大50%まで短縮できるため、結果がより速く得られます(図 "RT-PCR時間を顕著に短縮")。従って、サンプル数を大幅に増やしたり、他の実験者と1 台のサイクラーを効率的に共有できます。別々の反応ではなく同一反応内でコントロール遺伝子と標的遺伝子を増幅することで、マニュアルでの作業によるエラーが最小限に抑えられ、遺伝子定量の信頼性が増大します(図"信頼性のある相対定量")。QuantiFast Multiplex RT-PCR Kitに添付の特化されたマスターミックスにより、迅速なマルチプレックス反応のセットアップができ、singleplex RT-PCRデータに匹敵するマルチプレックスRT-PCR データを提供し、初めての実験でも良好な結果が得られます(図 "triplex とsingleplex RT-PCR で同等の結果を実現")。
QuantiFast Multiplex RT-PCR Kit を用いると、ターゲット量のわずかな差異を明確に判別できます。本キットは、2 倍希釈しかしていないテンプレートを用いた場合でも、テンプレート中で量が大きく異なるターゲットを正確に定量できます。最高4 種類のターゲットのマルチプレックスリアルタイムPCR が性能を損なうことなく高速で行なえます(図 "卓越した感度の4-plex RT-PCR")。
Fundamento
QuantiFast Multiplex RT-PCR Kitsでは最適化実験の必要はなく、標準サイクラーまたは高速サイクラーのどちらでも幅広いダイナミックレンジの高感度かつ迅速な結果が得られます(フローチャート "QIAGEN multiplex kits")。特別に開発された高速PCR 用バッファーは斬新なQ-Bondを含有し、変性、アニーリングおよびエクステンション時間を顕著に短縮します(図"プライマーの高速なアニーリング")。
別々の反応ではなく同一反応内でコントロール遺伝子と標的遺伝子を増幅することで、マニュアルでの作業によるエラーが最小限に抑えられ、遺伝子定量の信頼性が高くなります。QuantiFast Multiplex RT-PCR Buffer のK+ およびNH4+ イオン配合比により、プライマーの特異的なアニーリングを促進され、さらにユニークなFactor MP が特異的に結合したプライマーを安定化します(図 "ユニークなPCRバッファー")。さらに、至適化済み逆転写酵素ミックスにより、わずか20分で効率的なにcDNAを合成できます。また、HotStarTaq Plus DNA Polymeraseは厳密なホットスタートが可能で、非特異的産物の形成を防ぎます。
| 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 |
| QuantiFast Multiplex RT-PCR Buffer |
NH4+ イオンおよび 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PCR結果 |
| 合成添加剤Factor MP |
同一チューブで4遺伝子まで信頼できるマルチプレックス解析が可能 |
| ユニークなQ-Bond を含む |
PCR反応時間が短縮されるため迅速に結果が得られ、1日あたりの反応数を増やせる |
| ROX色素† |
Applied Biosystems製装置およびAgilent製装置での蛍光シグナルを補正 |
ROXが必要なサイクラーで正確な定量。他のリアルタイムサイクラーでの反応を妨害しない |
| QuantiFast RT Mix |
RNAへのアフィニティーが高い逆転写酵素の特殊な配合 |
複雑な二次構造のRNAでも、わずか20分で転写可能 |
Procedimento
QuantiFast Multiplex RT-PCR Kit は反応条件やサイクリング条件の至適化が不要で即使用可能なマスターミックスです。テンプレートRNA、プライマープローブセットをマスターミックスに加えハンドブックのプロトコール通りに操作するだけで、どのリアルタイムサイクラーでも迅速に信頼性の高い結果が得られます。マスターミックス中にROX passive reference dye が入ったキットまたは入っていないキットをお求めいただけますので、実質的にあらゆるリアルタイムサイクラーで使用できます(表を参照)。ROX濃度が至適化されているため、コピー数が少ない場合の検出でも自動データ解析を行なえます。
| マスターミックスに添加済み |
QuantiFast Multiplex RT-PCR Kit |
Applied Biosystems 7500以外のApplied Biosystemsの全てのサイクラー |
| 別チューブで添付 |
QuantiFast Multiplex RT-PCR +R Kit |
Applied Biosystems 7500 および
Bio-Rad、Cepheid、Eppendorf、QIAGEN、Roche、Agilent、その他の会社のサイクラー |
Aplicações
QuantiFast Multiplex RT-PCR Kit はほとんどのリアルタイム用サーマルサイクラーで遺伝子発現解析、その他のアプリケーションに使用可能です。これには、Applied Biosystems およびBio-Rad、Cepheid、Eppendorf、Roche、Agilent 製サイクラーを含みます。Rotor-Gene Q および他のRotor-Gene サイクラーを使用する場合は、これらの高速サイクリング用に開発されたRotor-Gene Multiplex RT-PCR Kitの使用をお勧めします。
|
Característica
|
Especificações
|
|
Applications
|
Real-time quantification of RNA targets in a multiplex format
|
|
Reaction type
|
Real-time one-step RT-PCR
|
|
Real-time or endpoint
|
Real-time
|
|
Sample/target type
|
RNA
|
|
Single or multiplex
|
Multiplex
|
|
SYBR Green I or sequence-specific probes
|
Sequence-specific probes
|
|
Thermal cycler
|
Real-time cyclers dedicated for multiplex PCR (e.g., most Applied Biosystems real-time PCR cyclers, Roche LightCycler 480, and Bio-Rad iCycler iQ)
|
|
With or without ROX
|
Available with ROX in master mix or with ROX as separate vial
|
|
|
|
|
For quantitative, multiplex, real-time one-step RT-PCR with fast cycling using sequence-specific probes
|
Mostrar detalhes
|
Imagens
Significantly reduced RT-PCR times.
QuantiFast Multiplex Kits reduce total RT-PCR run time by up to 50% in real-time one-step RT-PCR (40 cycles run; comparison with QuantiTect Multiplex Kits). I: iCycler iQ; L1: LightCycler 480; L2: LightCycler 2.0; A1: ABI PRISM 7900; A2: Applied Biosystems 7500 Fast System; M: Mx3005P.
Reliable relative quantification.
Cell line X was treated with one of two compounds (Y or Z) or left untreated (U). RNA was purified and duplex, real-time one-step RT-PCR was carried out on the Applied Biosystems 7500 Fast System using the QuantiFast Multiplex RT-PCR +R Kit and TaqMan Gene Expression Assays for myogenin and GAPDH. For each treatment, 5 independent experiments were performed. [A] Changes in myogenin expression were detected with high accuracy and reproducibility (green curves). The expression of the housekeeping gene GAPDH was similar in all experiments (blue curves), allowing normalization of myogenin expression levels using the ΔΔCT method of relative quantification. [B] The fold changes in normalized myogenin expression level relative to that in untreated cells were consistent between experiments, as indicated by the small error bars. (Data kindly provided by Angelika Meyer, Novartis Pharma AG, Basel, Switzerland).
Comparable results in triplex and singleplex RT-PCR.
Triplex and singleplex, real-time one-step RT-PCR were carried out on the LightCycler 480 using the QuantiFast Multiplex RT-PCR +R Kit and self-designed TaqMan assays for [A] RPS27A (a ribosomal protein), [B] GAPDH (a housekeeping gene), and [C] UBC (a housekeeping gene). The template was Ramos cell line RNA (10 ng, 1 ng, or 0.1 ng), and reactions were run in duplicate. The comparable CT values for triplex PCR (colored curves) and singleplex PCR (gray curves) and [D] the high PCR efficiencies demonstrate the reliability of triplex PCR with the QuantiFast Multiplex RT-PCR +R Kit when analyzing targets of differing abundance.
Uncompromised sensitivity in 4-plex RT-PCR.
4-plex, real-time one-step RT-PCR was carried out on the Mx3005P using the QuantiFast Multiplex RT-PCR +R Kit and Primer Express designed TaqMan assays. Duplicate reactions were run using 10 ng RNA from Ramos cells as template. All 4 targets, which varied greatly in abundance, were reproducibly detected.
Fast primer annealing.
[A] Q-Bond in QuantiFast Buffer allows the DNA polymerase and primer to bind as a single complex, reducing the 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 annealing time.
QIAGEN multiplex kits.
QuantiFast Multiplex RT-PCR Kits provide a simple procedure for quantitative, multiplex, real-time PCR. In contrast to current methods, the kits eliminate the need for optimization of the concentrations of primers, Mg2+, and Taq DNA polymerase, even for difficult assays (e.g., assays in which the copy number of the target gene is much smaller than that for the reference gene).
Unique buffer promotes stable and efficient primer annealing.
[A] NH4+ ions prevent nonspecific primers from annealing to the template. [B] Synthetic Factor MP, an innovative PCR additive, increases the local concentration of primers at the template. Together with K+ and other cations, Factor MP stabilizes specifically bound primers, allowing efficient primer extension by HotStarTaq Plus DNA Polymerase.
|