QuantiFast Probe PCR Kits

使用序列特异性探针进行快速两步法RT-PCR,适用于基因表达分析

Products

QuantiFast Probe PCR Kits 旨在用于分子生物学应用。这些产品不能用于疾病诊断、预防和治疗。
Image

QuantiFast Probe PCR Kit (2000)

Cat. No. / ID. / ID.   204256

IMPORTANT NOTE: As announced earlier, the production of the QuantiFast kits has been discontinued since mid-2021. Hence, these products will be available only until stocks last.   For 2000 x 25 µl reactions: 25 ml 2x QuantiFast Probe PCR Master Mix (contains ROX dye), 20 ml RNase-Free Water
This kit is being phased out. We recommend switching to the QuantiNova successor product. For more information and FAQs on this transition, visit: www.qiagen.com/PCRresource.

特点

  • 快速获得结果,节约多达60%的时间
  • 灵敏的检测低拷贝模板
  • 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的PCR仪

产品详情

QuantiFast Probe PCR Kits利用序列特异性探针,可对cDNA进行快速、灵敏的两步法real-time PCR定量检测。Q-bond技术和优化的预混液使real-time RT-PCR的运行时间缩短,不仅在快速PCR仪上,即使在普通PCR仪上也可快速完成反应。即用型预混液含有热启动酶和独特的PCR缓冲液,确保在任何real-time PCR仪上无需优化即可进行灵敏的定量PCR检测。提供两种试剂盒规格。对于需要ROX进行荧光校准的PCR仪,可选择QuantiFast Probe PCR Kit。对于其他PCR仪,可选择QuantiFast Probe PCR +ROX Vial Kit。为了便于使用,预混液可保存在2–8°C。

绩效

QuantiFast Probe PCR Kits可获得高灵敏度结果,优于其它real-time PCR试剂盒(参见"Sensitive two-step RT-PCR on the iCycler iQ"和" Sensitive two-step RT-PCR")。 可缩短60%的PCR运行时间(参见" Significantly reduced PCR times"),更快获得结果,不影响PCR反应效果(参见" Faster results without compromising sensitivity")。可以明显提高您的样品通量,您也可以同其他用户共用PCR仪。

Sensitive two-step RT-PCR on the iCycler iQ
平均CT
人类白血球cDNA (ng) QuantiFast Probe PCR +ROX Vial Kit BV供应商的试剂盒
100 29.13 31.13
10 32.50 34.43
1 36.15 40.00
无模板对照 40.00 40.00
Triplicate reactions were run using a Primer Express designed TaqMan assay for IL8 (a chemokine). The QuantiFast Kit provided lower CT values than the kit from Supplier BV (which was used according to a fast-cycling protocol), allowing transcript quantification from just 1 ng cDNA.

QuantiFast Probe PCR Kits可在大范围内进行准确的定量检测(参见" Wide dynamic range and high sensitivity")。可靠检测稀释倍数高达7个数量级的模板稀释液(参见"Detection over 7 logs of template in two-step RT-PCR on the LightCycler 2.0 "和"Detection over 7 logs of template")。

Detection over 7 logs of template in two-step RT-PCR on the LightCycler 2.0
平均CT
人类白血球cDNA QuantiFast Probe PCR +ROX Vial Kit R供应商的试剂盒
100 ng 11.99 14.56
10 ng 15.54 17.99
1 ng 19.22 22.16
100 pg 22.63 25.14
10 pg 26.43 30.15
1 pg 30.02 34.43
100 fg 33.42 37.10
无模板对照 40 40.00
PCR效率 90% 80.1%
Duplicate reactions were run using a set of primers and FRET probes for b-2 microglobulin from supplier R. The QuantiFast Kit provided lower CT values and higher PCR efficiency that the kit from Supplier R (which was used according to the fast cycling protocol ).
查看图表

原理

QuantiFast Probe PCR Kits可在大范围内进行快速、灵敏的定量检测,适用于标准和快速PCR仪。该试剂盒专为各种序列特异性探针设计,包括水解探针(TaqMan探针和其他双标记探针)和FRET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")。缓冲液中K+和NH4+离子的平衡配比可促进特异性的引物退火,确保高度灵敏和特异的RT-PCR反应(参见" Specific primer annealing")。此外,HotStarTaq Plus DNA Polymerase需要严格的热启动,可避免生成非特异性产物。

2x QuantiFast Probe PCR Kit的成分*
成分 特点 优势
HotStarTaq Plus DNA Polymerase 95ºC加热3分钟活化 在室温进行qPCR反应体系构建
QuantiFast Probe 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
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)。 ROX染料或者在预混液中,或者单独提供。
查看图表

程序

QuantiFast Probe PCR Kits包含即用型预混液,无需对反应和扩增条件进行优化。只需按照操作手册进行实验,即可快速获得可靠的结果。提供含有和不含ROX染料的预混液(参见下表)。由于ROX浓度经优化,即使是低拷贝数也可通过数据自动分析进行检测。

Choosing the right QuantiFast Probe PCR Kit
ROX染料试剂盒 适用的PCR仪
混合在预混液中 QuantiFast Probe PCR Kit 除Applied Biosystems 7500外的所有Applied Biosystems仪器
单独装在管中 QuantiFast Probe PCR +ROX Vial Kit Applied Biosystems 7500、Agilent、Bio-Rad、Cepheid、QIAGEN、Eppendorf、Roche和其他供应商的PCR仪

为获得最佳的两步法real-time RT-PCR结果,我们推荐应用QuantiTect Reverse Transcription Kit合成cDNA。该试剂盒可在20分钟内快速合成cDNA,同时去除基因组DNA。

QuantiFast Probe Assays是预制的适合全基因组的试剂,使用水解探针法检测;包含在QuantiFast Probe PCR Kit中,可确保在两步法qRT-PCR中获得可靠结果。

应用

QuantiFast Probe PCR Kits可在任何real-time PCR仪上利用序列特异性探针进行cDNA基因表达分析。适用于Applied Biosystems、Bio-Rad、Cepheid、Eppendorf、Roche和Agilent的PCR仪。在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 Probe PCR Kit。

辅助数据和图表

规格

特点规格
applicationsProbe-based, real-time PCR, two-step RT-PCR
sampletargettypecDNA, DNA
realtimeorendpointReal-time
reactiontypeReal-time and two-step RT-PCR
singleormultiplexSingle
withorwithoutroxAvailable with ROX in master mix and with ROX as separate vial
sybrgreeniorsequencespecificprobesSequence-specific probes
thermalcyclerAll real-time cyclers (e.g. Roche LightCycler®, Corbett Rotor-Gene, Applied Biosystems)

资源

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
快速启动实验方案 (2)
试剂盒操作手册 (1)
For fast, quantitative, real-time PCR and two-step RT-PCR using sequence-specific probes
Certificates of Analysis (1)