QuantiFast SYBR^® Green PCR Kit

SYBR Greenを用いた高速リアルタイムPCRおよび2ステップqRT-PCR用

低コピーターゲットでも、特異的で高感度な検出
至適化済みマスターミックスで至適化せずに正確な結果を実現
5分間の酵素活性化と高速サイクリング条件
幅広いテンプレート量を正確に検出
標準的および高速サイクラーに対応する共通プロトコール

QuantiFast SYBR Green PCR Kitは、SYBR Green I検出を用いたリアルタイムPCRまたは2ステップRT-PCRで迅速かつ特異的にgDNAまたはcDNAターゲットを定量します。Q-bondテクノロジーと至適化済みで即使用可能な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標準的なサイクラーでも、より短時間でリアルタイムPCRを実現します。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、至適化の必要はなく、全てのリアルタイムサイクラーで高感度な定量PCR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。

Ordering Information
Product Details
Specifications
Product Resources

Please log in Show cart

Product	Cat. no.	List price:
QuantiFast SYBR Green PCR Kit (400) For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast SYBR Green PCR Master Mix (contains ROX dye), 2 x 2 ml RNase-Free Water	204054	4.530,00 kr	Add to cart
QuantiFast SYBR Green PCR Kit (2000) For 2000 x 25 µl reactions: 25 ml 2x QuantiFast SYBR Green PCR Master Mix (contains ROX dye), 20 ml RNase-Free Water	204056	18.385,00 kr	Add to cart
QuantiFast SYBR Green PCR Kit (4000) For 4000 x 25 µl reactions: 2 x 25 ml 2x QuantiFast SYBR Green PCR Master Mix (contains ROX dye), 20 ml RNase-Free Water	204057	25.345,00 kr	Add to cart

Please log in Show cart

The QuantiFast SYBR^® Green PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Main Image Navi

The QuantiFast SYBR^® Green PCR Kit reduces total PCR run time by up to 60% in real-time two-step RT-PCR on standard cyclers (40 cycles without melting curve analysis; comparison with standard QIAGEN real-time PCR kits). L: LightCycler 2.0; A1: ABI PRISM 7900; A2: Applied Biosystems 7500; A3: ABI PRISM 7000.|The QuantiFast SYBR^® Green PCR Kit was used to detect the Y-chromosome-specific single-copy gene SRY in genomic DNA from a male donor using the Mastercycler ep realplex. [A] Curves for 1000 down to 1.5 copies can be clearly distinguished from each other. [B] A plot of copy number (log) versus C_T value demonstrates high linearity.|Expression of MYC (a proto-oncogene) in human leukocytes was analyzed by real-time two-step RT-PCR on the Applied Biosystems 7500 Fast System. Duplicate reactions were run using 10-fold cDNA dilutions (10 ng to 10 pg). [A] Fast cycling mode with the QuantiFast Kit gave similar C_T values to [B] standard cycling mode with the QuantiTect Kit. In contrast, a kit for standard cycling from Supplier A_II gave worse C_T values not only in [C] fast cycling mode, but also in [D] recommended standard cycling mode.|Expression of MYC (a proto-oncogene) in human leukocytes was analyzed using the iCycler iQ and [A] the QuantiFast SYBR^® Green PCR Kit or [B] a kit from Supplier B_v. Duplicate reactions were run using 10-fold cDNA dilutions (100 ng to 0.1 ng). The instrument-dedicated kit from Supplier B_V was used according to the fast cycling protocol. The QuantiFast Kit gave lower C_T values, indicating greater sensitivity.|A balanced combination of KCl and (NH₄)₂SO₄ promotes specific annealing of primers and probes to the PCR template. K⁺ binds to phosphate groups on double-stranded DNA, stabilizing annealing of primers and probes. NH₄⁺ destabilizes weak hydrogen bonds between mismatched bases.|[A] Q-Bond in QuantiFast Buffer 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

Fastサイクリングモードで使用した他のリアルタイムPCRキットに比較して、QuantiFast SYBR Green PCR Kitでは卓越した特異性と感度の高い結果が得られます（図"高感度な2ステップRT-PCR"）。PCR時間が最大60％短縮され（図"PCR時間を顕著に短縮"）、PCRの性能を妥協しなくても結果をより速く得られます（図"感度を損なわずにより迅速な結果を実現"）。従って、サンプル処理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。

QuantiFast SYBR Green PCR Kit は、数段階に対数希釈したテンプレート液で正確な定量を実現します。低コピー数のターゲットのわずかな差異も明確に区別することができます（図"わずかなコピー数の差異を解析"）。

Principle

QuantiFast SYBR Green PCR Kitでは、標準的なサイクラー、高速サイクラーともに、広範なダイナミックレンジで非常に高感度かつ特異的な結果が得られます。マスターミックスの蛍光色素SYBR Green Iでは多種類のターゲットの解析が可能で、ターゲット特異的な標識プローブを合成する必要もありません。新規PCR添加剤のQ-Bondを含有する特別に開発された高速PCR用バッファーにより、変性、アニーリングおよびエクステンション時間も顕著に短縮されます（図"プライマーの高速なアニーリング"）。PCRバッファー中のK⁺およびNH₄⁺の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が促進され、高いPCR効率と感度を実現します（図"特異的なプライマーアニーリング"）。さらに、HotStarTaq Plus DNA Polymeraseで活性化に必要な時間は95℃ではわずか5分で、厳密なホットスタートを行なえるため、非特異的な産物の形成を抑えます。

2x QuantiFast SYBR Green PCR Kitの成分*
成分	特長	利点
HotStarTaq Plus DNA Polymerase	95℃、5分間の活性化	室温での定量PCRのセットアップ
QuantiFast SYBR Green PCR Buffer	NH₄⁺/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
QuantiFast SYBR Green PCR Buffer	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
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	高感度な定量
ROX色素	Applied Biosystems社およびオプションのAgilent社の装置で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

Procedure

QuantiFast SYBR Green 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。プライマーおよびDNAテンプレートを即使用可能なPCRマスターミックスに添加するだけで、反応を開始することができます。ハンドブックに記載されているプロトコールに従うだけで、全てのリアルタイムサイクラーで迅速で正確な結果が得られます。

2ステップリアルタイムRT-PCRで最適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。本キットは、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。

SYBR Greenを用いた遺伝子発現解析にはQuantiTect Primer Assayをお薦めします。QuantiTect Primer Assayは、ヒト、マウス、ラット、その他の生物種の全て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 Webポータルサイトで簡単にオーダーできます。

Applications

QuantiFast SYBR Green PCR KitはcDNAターゲットの遺伝子発現解析および定量gDNA解析に使用できます。QuantiFast SYBR Green PCR KitはApplied Biosystems、Bio-Rad、Cepheid、Eppendorf、RocheおよびAgilent社製などの市販されている全てのリアルタイムサイクラーに対応します。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特別に開発されたRotor-Gene SYBR Green PCR Kitの使用をお勧めします。

Feature	Specifications
Applications	SYBR Green-based, real-time PCR, two-step RT-PCR
Reaction type	Real-time and two-step RT-PCR
Real-time or endpoint	Real-time
Sample/target type	cDNA, DNA
Single or multiplex	Single
SYBR Green I or sequence-specific probes	SYBR Green I
Thermal cycler	Applied Biosystems, Bio-Rad, Cepheid, QIAGEN, Eppendorf, Roche, and Agilent
With or without ROX	With ROX
With/without hotstart	With hotstart

Brochures & Guides

Sort options Sort alphabetically
	Flexible RNAi Technologies You Can Rely On - (EN)	Show details
	(EN) - Analyzing Gene Expression and Regulation	Show details
	Critical Factors for Successful Real-Time PCR - (EN)	Show details

FAQs

Sort options Sort alphabetically
	What should I use as a standard for absolute quantification in real-time PCR? FAQ ID -1085	View
	What is a QuantiTect Primer Assay? FAQ ID -1141	View
	How many reactions can I perform with the new QuantiFast Kits for real-time PCR? FAQ ID -1425	View
	Can I adjust the ROX concentration in the QuantiFast master mix? FAQ ID -1427	View
	Can QuantiFast PCR Kits be used on real-time PCR instruments without fast cycling options? FAQ ID -1428	View
	Do you offer trial-kit sizes for the new QuantiFast Kits? FAQ ID -1429	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
	Does the master mix in the QuantiFast Kits contain dUTP to allow UNG treatments? FAQ ID -1431	View
	What is the detection limit of the QuantiFast Kits for real-time PCR? FAQ ID -1432	View
	What QuantiFast Kit should be used on the Eppendorf Mastercycler ep realplex? FAQ ID -1436	View
	Do special settings have to be used for QuantiFast PCR Kits on the Eppendorf Mastercycler ep realplex? FAQ ID -1437	View
	How much time will be saved when switching from standard cycling to fast cycling with QuantiFast Kits? FAQ ID -1438	View
	Do the QuantiTect Primer Assays work with QuantiFast SYBR Green Kits? FAQ ID -1439	View
	Can 2 µl reaction volumes be used with QuantiFast PCR Kits? FAQ ID -1440	View
	How do QuantiFast PCR Kits compare to QuantiTect PCR Kits for quantitative real-time PCR? FAQ ID -1441	View
	Why is the QuantiFast denaturation step different for PCR and RT-PCR runs in the two-step protocol for the ABI 7500 and other cyclers? FAQ ID -1442	View
	Why do QuantiFast SYBR Green PCR Kits require such a high primer concentration? FAQ ID -1443	View
	Does the high primer concentration required by QuantiFast SYBR Green PCR Kits impair annealing specificity? FAQ ID -1444	View
	Why is the storage time for QuantiFast PCR Kits shorter than that for QuantiTect PCR Kits? FAQ ID -1446	View
	Why is the reaction volume for QuantiFast PCR Kits lower than that for QuantiTect PCR Kits? FAQ ID -1447	View
	Why is the activation time for HotStarTaq Plus Polymerase in the QuantiFast SYBR Green Kits different from that for QuantiFast Probe Kits? FAQ ID -1449	View
	Why is a 2-step (and not a 3-step) cycling protocol recommended for QuantiFast SYBR Green PCR Kits? FAQ ID -1450	View
	Will QuantiTect Primer Assays work at an annealing temperature of 60ºC with QuantiFast SYBR Green PCR and RT-PCR Kits? FAQ ID -1553	View
	What is Q-Bond used in the QIAGEN Fast Cycling PCR and QuantiFast Kits? FAQ ID -1554	View
	Which real-time PCR kits are recommended downstream of the QuantiTect Whole Transcriptome Kit? FAQ ID -1592	View
	Is the master mix of QuantiFast Kits for real-time PCR aliquoted into several tubes to prevent cross-contamination? FAQ ID -1697	View
	Have QuantiTect Primer Assays been tested with QuantiFast SYBR Green PCR Kits on the Mastercycler ep realplex? FAQ ID -1714	View
	QuantiTect Primer Assays are bioinformatically validated, genomewide primer sets. What does “bioinformatically validated” mean? FAQ ID -1982	View
	What are the main differences between Rotor-Gene and QuantiTect or QuantiFast PCR Kits? FAQ ID -2119	View
	Do QuantiTect Primer Assays also work with Rotor-Gene SYBR Green PCR Kits? FAQ ID -2123	View
	Will Uracil-N-Glycosylase (UNG) completely remove contaminating amplicons when using QuantiTect +UNG Kits? FAQ ID -2139	View
	Do you have any information or guidelines regarding the choice of reference genes for real-time PCR? FAQ ID -2371	View
	Is the QuantiFast SYBR Green PCR Kit compatible with the ViiA7 cycler from Applied Biosystems? FAQ ID -2652	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 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 delta Rn value? FAQ ID -2681	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
	Why do replicates in real-time PCR have different plateau heights? FAQ ID -539	View
	Why does my realtime PCR assay quality decrease over time? FAQ ID -589	View
	Do you have information on the use of recombinant DNA and RNA as absolute standards for realtime RT-PCR? FAQ ID -729	View
	Why should DNA or cDNA targets be less than 250 bp long for real-time PCR? FAQ ID-751	View
	What annealing temperature should be used with the QuantiTect Primer Assays? FAQ ID -849	View

Kit Handbooks

Sort options Sort alphabetically
	QuantiFast SYBR Green PCR Handbook - (EN) For fast, quantitative, real-time PCR and two-step RT-PCR using SYBR Green I	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

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

Scientific Posters

Sort options Sort alphabetically
	Quantitative RT-PCR analysis with genomewide, validated primer sets - (EN)	Show details
	(EN) - New concepts for accelerated real-time PCR analysis	Show details

Supplementary Protocols

Sort options Sort alphabetically
	ABI PRISM® 7000 Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	ABI PRISM® 7700 Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	ABI PRISM® 7900 Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	Applied Biosystems® 7500 Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	DNA Engine Opticon® Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	iCycler iQ® Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	LightCycler® 1.x Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	LightCycler® 2.0 Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	LightCycler® 480 Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	Mastercycler® ep realplex Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	Mx3005P® Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details
	Rotor-Gene™ Software Setup for the QuantiFast™ SYBR® Green PCR Kit - (EN)	Show details

Quick-Start Protocols

Sort options Sort alphabetically
	QuantiFast SYBR Green PCR Kit (EN)	Show details

Safety Data Sheets

Sort options Sort alphabetically
	Safety Data Sheets Download Safety Data Sheets for QIAGEN product components.	View

Safety Data Sheets

Sort options Sort alphabetically
	MSDS QuantiFast SYBR Green PCR Kit (400)
	MSDS QuantiFast SYBR Green PCR Kit (2000)
	MSDS QuantiFast SYBR Green PCR Kit (4000)
	CofA

Images

Significantly reduced PCR times.

Resolution of small differences in copy number.

The QuantiFast SYBR^® Green PCR Kit was used to detect the Y-chromosome-specific single-copy gene SRY in genomic DNA from a male donor using the Mastercycler ep realplex. [A] Curves for 1000 down to 1.5 copies can be clearly distinguished from each other. [B] A plot of copy number (log) versus C_T value demonstrates high linearity.

Faster results without compromising sensitivity.

Expression of MYC (a proto-oncogene) in human leukocytes was analyzed by real-time two-step RT-PCR on the Applied Biosystems 7500 Fast System. Duplicate reactions were run using 10-fold cDNA dilutions (10 ng to 10 pg). [A] Fast cycling mode with the QuantiFast Kit gave similar C_T values to [B] standard cycling mode with the QuantiTect Kit. In contrast, a kit for standard cycling from Supplier A_II gave worse C_T values not only in [C] fast cycling mode, but also in [D] recommended standard cycling mode.

Senstivite two-step RT-PCR on the iCycler iQ.

Sensitive two-step RT-PCR.

Expression of MYC (a proto-oncogene) in human leukocytes was analyzed using the iCycler iQ and [A] the QuantiFast SYBR^® Green PCR Kit or [B] a kit from Supplier B_v. Duplicate reactions were run using 10-fold cDNA dilutions (100 ng to 0.1 ng). The instrument-dedicated kit from Supplier B_V was used according to the fast cycling protocol. The QuantiFast Kit gave lower C_T values, indicating greater sensitivity.

Specific primer annealing.

A balanced combination of KCl and (NH₄)₂SO₄ promotes specific annealing of primers and probes to the PCR template. K⁺ binds to phosphate groups on double-stranded DNA, stabilizing annealing of primers and probes. NH₄⁺ destabilizes weak hydrogen bonds between mismatched bases.

Fast primer annealing.

[A] Q-Bond in QuantiFast Buffer 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.

QuantiFast SYBR® Green PCR Kit

QuantiFast SYBR^® Green PCR Kit