NRAS Pyro Kit

基于序列定量分析NRAS基因的突变

S_1084_5_GEN_V2

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NRAS Pyro Kit (24)

Cat. No. / ID:  970530

For 24 reactions: Sequencing Primers, PCR Primers, Unmethylated Control DNA, PyroMark PCR Master Mix, CoralLoad Concentrate, Buffers, and Reagents
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NRAS Pyro 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。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

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Features

  • 实时获得多个结果
  • 准确定量NRAS基因中的突变
  • 简便检测复杂突变
  • 序列信息提供检测的内置参照
  • 灵活的后续分析

Product Details

NRAS Pyro Kit是用于鉴定NRAS基因突变的分子检测试剂盒。该试剂盒包含扩增NRAS基因所需的引物和试剂、外加缓冲液、引物和试剂,在PyroMark Q24 System上使用焦磷酸测序技术实时定量检测突变。

Principle

NRAS Pyro Kit用于实时定量检测人NRAS基因编码子12、13和61的突变,在PyroMark Q24 System上使用焦磷酸测序技术检测。NRAS基因编码一种GTPase,该GTPase是成神经细胞瘤RAS病毒(v-ras)原癌基因同源物。在所有恶性黑素瘤中,有将近13–25%发生NRAS突变,通常是编码子12、13和61的突变。这些突变影响NRAS信号通路的活性。

可检测以下种类的突变:

  • 编码子12/13:G12A、G12C、G12D、G12R、G12S、G12V、G13A、G13C、G13D、G13R、G13S和G13V
  • 编码子61:Q61E、Q61H、Q61K、Q61L、Q61P和Q61R

Procedure

该试剂盒包含2部分实验:一个用于检测编码子12和13的突变;另一个用于检测编码子61的突变(参见" Illustration of the NRAS assay"和" Pyrogram trace of a normal genotype in codon 12–13")。运用PCR技术对这两个片段分别进行扩增,测定特定区域的核酸序列。使用该法进行定量和定性分析时,特定区域周围的核酸序列峰被视为标准峰和参照峰。

使用针对编码子12/13和编码子61的引物对两个片段PCR扩增后,将扩增子固定在高性能链霉素亲和琼脂糖上。将单链DNA和相应序列的引物退火处理得到DNA。然后在PyroMark Q24 System上设置程序和运行程序对样品进行分析。运行后可以通过调节"Sequence to Analyze"来检测稀有突变(参见" Pyrogram trace of a GGT to AGT mutation in base 1 of codon 12"和" Pyrogram trace of the reanalysis of the sample in the previous figure")。

See figures

Applications

NRAS Pyro Kit用于检测人NRAS基因编码子12、13和61的突变。该试剂盒确保准确的定量突变分析,适用于各种研究领域,包括癌症和表观遗传学研究。

可检测以下种类的突变:

  • 编码子12/13:G12A、G12C、G12D、G12R、G12S、G12V、G13A、G13C、G13D、G13R、G13S和G13V
  • 编码子61:Q61E、Q61H、Q61K、Q61L、Q61P和Q61R

Supporting data and figures

Resources

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
分析软件 (1)
RAS Extension Plug-in 1.3
SOFTWARE (1MB)
Version 1.3.0.7
For use with PyroMark Q24 Software version 2.0.8
Safety Data Sheets (1)
Certificates of Analysis (1)
Analysis Software (1)
RAS Extension Plug-in 1.3
SOFTWARE (1MB)
Version 1.3.0.7
For use with PyroMark Q24 Software version 2.0.8

FAQ

Can I use more than 10ng DNA for the PCR reaction for the Pyro kits?
The NRAS Pyro, EGFR Pyro. BRAF PyroKRAS Pyro, and UGT1A1 Pyro Kits all require that no more than 10ng of DNA be used for the PCR reaction.  Too much DNA can sometimes lead to inhibition problems in the PCR reaction.
FAQ ID -2379
Which purification kits are recommended for the Pyro kits?

The purification kits recommended for the BRAF Pyro, EGFR Pyro, KRAS Pyro, and NRAS Pyro kits are:

Paraffin-embedded tissue

Blood

FAQ ID -2380
What plug-ins are available for the Pyrosequencing kits?
Plug-Ins are available for:
  • EGFR Pyro Kit 
  • KRAS Pyro Kit
  • NRAS Pyro Kit
  • RAS extension Kit
  • BRAF Pyro Kit
  • These plug-ins are available for download on the respective catalog page under the Product Resources tab in the Analysis Software section. Note: PyroMark Q24 software version 2.0.7 is needed for the usage of the Plug-ins.
    FAQ ID -2381
    Can PyroMark Gold reagents be vortexed?
    Reconstiuted enzyme and substrate of PyroMark Gold Reagents, should not be vortexed since this could lead to conformational changes which affect the activity.
    FAQ ID -2844