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。

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
$577.00
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
$2,312.00
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
$3,210.00
Add to 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.
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Significantly reduced PCR times.|Resolution of small differences in copy number.|Faster results without compromising sensitivity.|Sensitive two-step RT-PCR.|Specific primer annealing.|Fast primer annealing.|
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 CT 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 CT values to [B] standard cycling mode with the QuantiTect Kit. In contrast, a kit for standard cycling from Supplier AII gave worse CT 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 Bv. Duplicate reactions were run using 10-fold cDNA dilutions (100 ng to 0.1 ng). The instrument-dedicated kit from Supplier BV was used according to the fast cycling protocol. The QuantiFast Kit gave lower CT 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
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
* 也含有dNTP混合液(dATP、dCTP、dGTP和dTTP)。

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。

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Kit Handbooks
2
For fast, quantitative, real-time PCR and two-step RT-PCR using SYBR Green I
Show details
For genomewide, ready-to-use real-time RT-PCR assays using SYBR Green detection
Show details
Safety Data Sheets
1
Download Safety Data Sheets for QIAGEN product components.
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Images
Significantly reduced PCR times
Significantly reduced PCR times.
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.
Resolution of small differences in copy number on the Mastercycler ep realplex.
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 CT value demonstrates high linearity.
Faster results without compromising sensitivity on the Applied Biosystems 7500 Fast System.
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 CT values to [B] standard cycling mode with the QuantiTect Kit. In contrast, a kit for standard cycling from Supplier AII gave worse CT 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 Bv. Duplicate reactions were run using 10-fold cDNA dilutions (100 ng to 0.1 ng). The instrument-dedicated kit from Supplier BV was used according to the fast cycling protocol. The QuantiFast Kit gave lower CT values, indicating greater sensitivity.
Specific primer annealing using a balanced combination of K+ and NH4+ ions
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 with Q-Bond
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.