QuantiFast SYBR^® Green PCR Kit

使用SYBR Green I的快速两步法定量RT-PCR，适用于基因表达分析

快速获得结果，可节省多达60%的时间
可特异性检测低拷贝数的模板
准确检测各种起始量的模板
优化的即用型预混液用于快速PCR反应
通用的操作流程适用于标准和快速PCR仪

QuantiFast SYBR Green PCR Kit应用SYBR Green I法两步法real-time RT-PCR，可对目标cDNA进行快速、特异性的定量检测。Q-Bond和优化的预混液可缩短real-time PCR反应时间，适用于标准或快速PCR仪。即用型预混液中的热启动酶和独特的PCR缓冲液可确保在所有real-time PCR仪上进行灵敏的qPCR，无需优化。为便于使用，预混液可保存在2–8°C。

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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	S Fr. 702.00	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	S Fr. 2'750.00	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	S Fr. 3'851.00	Add to cart

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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.

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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

QuantiFast SYBR Green PCR Kits通过快速PCR反应，实现了特异性、灵敏的real-time PCR检测（参见"Sensitive two-step RT-PCR"）。PCR反应时间缩短60%（参见"Significantly reduced PCR times"），是您更快获得结果，而不影响PCR效果（参见"Faster results without compromising sensitivity"）。您也可以大幅度提高样本通量，或与其他用户高效的共用PCR仪。

QuantiFast SYBR Green PCR Kit可清楚区分模板量的细微差别。即使低拷贝数的模板只有细微差别，该试剂盒也能准确定量（参见"Resolution of small differences in copy number"）。

Principle

QuantiFast SYBR Green PCR Kits可在大范围内进行特异性、灵敏的检测，适用于标准和快速PCR仪。预混液中的SYBR Green I染料可分析多个目标核酸，无需合成序列特异性探针。特制的快速PCR缓冲液包含新型Q-Bond成分，可大大缩短变性、退火和延伸时间（参见"Fast primer annealing"）。PCR缓冲液中平衡的K⁺和NH₄⁺离子的配比可促进特异性的引物退火，确保高度灵敏和特异的PCR反应（参见"Specific primer annealing"）。此外，HotStarTaq Plus DNA Polymerase必须要在95°C加热5分钟才能活化，需要严格的热启动，可避免生成非特异性产物。

2x QuantiFast SYBR Green PCR Kit的成分*
成分	特点	优势
HotStarTaq Plus DNA Polymerase	95ºC加热5分钟活化	在室温进行qPCR反应体系构建
QuantiFast SYBR Green PCR Buffer	平衡的NH₄⁺和K⁺离子配比	引物探针的特异性结合确保获得可靠结果
QuantiFast SYBR Green PCR Buffer	独特的Q-Bond添加剂	PCR运行时间缩短，更快获得结果，一天内可完成更多PCR反应
SYBR Green I染料	与DNA双链结合时产生强荧光信号	高灵敏度扩增
ROX染料	对Applied Biosystems和Agilent PCR仪进行荧光信号的校准	对需要ROX染料的PCR仪进行校准，不影响PCR反应结果

Procedure

QuantiFast SYBR Green PCR Kit含有即用型预混液，无需对反应和扩增条件进行优化。只需在即用型PCR预混液中加入引物和cDNA模板，即可开始反应。按照操作手册中的操作流程，即可快速获得可靠的结果，适用于各种real-time PCR仪。

为获得最佳的两步法real-time RT-PCR结果，我们建议使用QuantiTect Reverse Transcription Kit合成cDNA。该试剂盒可在20分钟进行快速cDNA合成，并去除基因组DNA污染。

我们推荐适用QuantiTect Primer Assays进行SYBR Green法的基因表达分析。 QuantiTect Primer Assays是经生物信息学验证的引物，适用于人类、大鼠、小鼠和其他多个物种的全基因组。可直接在GeneGlobe订购该产品。

Applications

QuantiFast SYBR Green PCR Kits可用于cDNA的基因表达分析，适用于各种real-time PCR仪，包括Applied Biosystems、Bio-Rad、Cepheid、Eppendorf、Roche和Agilent的PCR仪。Rotor-Gene Q实时荧光定量PCR分析仪或其他Rotor-Gene PCR仪上使用时，我们推荐使用专为快速PCR研发的Rotor-Gene SYBR Green PCR Kit。

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

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	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

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	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

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	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

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

Scientific Posters

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	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

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	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

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	QuantiFast 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 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