Rotor-Gene SYBR® Green PCR Demo Kit

用于评估Rotor-Gene Q实时荧光定量PCR分析仪的性能

S_1517_GEF_PCR0030

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的(再)订购

Rotor-Gene SYBR Green PCR Demo Kit (80)

Cat. No. / ID:   204001

For 80 reactions: 2x Rotor-Gene SYBR Green PCR Master Mix, 10x QuantiTect Primer Assay for GPER, Standards, Unknown Samples, Buffer TE, RNase-Free Water
Rotor-Gene SYBR® Green PCR Demo Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的(再)订购

特点

  • 配合Rotor-Gene预混液评估Rotor-Gene Q实时荧光定量PCR分析仪的性能
  • 确定Rotor-Gene Q实时荧光定量PCR分析仪的精确度和可重复性
  • 包含所有组分的完整系统

产品详情

Rotor-Gene SYBR Green PCR Demo Kit配合Rotor-Gene预混液,用于展现Rotor-Gene Q实时荧光定量PCR分析仪的性能。试剂盒包含所有需要的反应组分,来通过SYBR Green-based real-time PCR对基因组DNA靶标进行定量。这包括预稀释的基因组DNA标准物、2个未知样本、优化的预混液和人G蛋白耦联雌激素受体1(GPER)基因的特异性引物对。使用此试剂盒可评估Rotor-Gene Q实时荧光定量PCR分析仪进行基因定量时的可靠性、可重复性和灵敏度。

原理

Rotor-Gene Kits配合Rotor-Gene Q实时荧光定量PCR分析仪可完成高精度real-time PCR。Rotor-Gene Kits中提供的即用型预混液可快速、准确的进行基因定量,无需反应优化,而Rotor-Gene Q实时荧光定量PCR分析仪采用独特的离心转子设计。PCR管放置到转子中,离心的PCR管穿过一道激发光源和一个流动空气腔内的检测器。这意味着管间几乎没有光照和温度变化,使real-time PCR定量分析具有高精度。此外,由于转子离心速度持续在400 rpm,使高速数据采集成为可能。

使用Rotor-Gene SYBR Green PCR Demo Kit可确保高度特异性的扩增,离子的平衡组合可减少非特异性引物退火(参见" Specific primer annealing")。使用Q-Bond可进行快速循环,不影响性能,Q-Bond是一种新颖的PCR添加剂,使PCR运行时间可少至45分钟(参见" Fast primer annealing")。

查看图表

程序

使用Rotor-Gene SYBR Green PCR Demo Kit进行SYBR Green法real-time PCR,可定量不同的基因组DNA靶标的拷贝数。每个反应包含:

  • 特定拷贝数的人基因组DNA模板
  • Rotor-Gene SYBR Green PCR Master Mix
  • 特异性分析人G蛋白耦联雌激素受体1(GPER)基因的QuantiTect Primer Assay

Rotor-Gene SYBR Green PCR Demo Kit手册包含3个实验方案。Rotor-Gene用户可选用手工反应体系构建的实验方案或使用QIAgility自动化反应体系构建的实验方案。反应体系构建后,用户可选择在Rotor-Gene Q实时荧光定量PCR分析仪上进行real-time PCR的实验方案。

使用QIAgility可避免手工移液步骤,紧凑的台式仪器可进行快速、高精度的PCR反应体系构建,可减少或消除由于人为失误造成的反应体系构建错误。QIAgility可配合Rotor-Gene Q实时荧光定量PCR分析仪和Rotor-Gene Kits使用,可简单分装液体到单管、联管和Rotor-Discs。

通过一套标准物(2000、1000、500、250和125个拷贝;每个标准物有4个平行分析)的CT值可产生一条标准曲线。标准曲线用来确定2个未知样本(500和250个拷贝;每个未知样本有24个重复[当手工反应体系构建时,每个未知样本可最少有4个重复])的拷贝数。此外,还进行4个无模板对照(NTC)反应。因此,在Rotor-Gene Q实时荧光定量PCR分析仪上有72个反应同时进行。

应用

Rotor-Gene SYBR Green PCR Demo Kit用于评估Rotor-Gene Kits配合Rotor-Gene Q实时荧光定量PCR分析仪的性能。此试剂盒与Rotor-Gene 3000或Rotor-Gene 6000兼容。使用此试剂盒可评估Rotor-Gene Q实时荧光定量PCR分析仪在基因定量时的精确度和可重复性。

辅助数据和图表

资源

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (1)
For evaluation of the performance of the Rotor-Gene Q
实验方案软件 (2)
Safety Data Sheets (1)
Certificates of Analysis (1)

FAQ

How do I create a workspace that is free of DNA contamination, prior to carrying out a qPCR experiment?

Any DNA contamination will artificially inflate the SYBR Green signal, yielding skewed gene expression profiles and false-positive signals. The most common source of DNA contamination is from PCR products generated during previous experiments. Such contamination is most often due to the improper disposal of tubes, tips, and gels that previously came into contact with PCR products. Additionally, PCR products may also contaminate pipettors, racks, work pads, and commonly used reagents such as water and buffers. To minimize the risk of contaminating your experiment with extraneous DNA, the following steps should be taken:

 

  • Remove a single aliquot of water from your PCR-grade stock, sufficient to complete the experiment. This minimizes the number of times that the stock container is opened, thereby minimizing contamination risks.
  • Use only fresh PCR-grade reagents and disposable labware.
  • Treat any labware (tubes, tips, and tip boxes) used in PCR with 10% bleach, before discarding.
  • Maintain a dedicated workspace for PCR setup (perhaps a PCR-only hood), away from areas of the lab where post-PCR work is done, such as running gels, enzyme digestions, and cloning.
  • Change the lab bench pads/papers often and decontaminate lab benches and labware (racks, pipettors, etc.) before each use by washing with 10% bleach, and/or exposing to UV light for at least 10 minutes. This serves to degrade and/or inactivate contaminating DNA.
  • Before, during, and after the experiment, minimize the opening and closing of any tubes or plates used during the experiment.  
FAQ ID -2654