KRAS Pyro Kit

基于序列定量分析KRAS基因中的突变

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KRAS Pyro Kit (24)

Cat. No. / ID: 970460

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

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KRAS 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。

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特点

实时获得多个结果
准确定量KRAS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

产品详情

KRAS Pyro Kit是用于鉴定KRAS基因中突变的分子检测试剂盒。该试剂盒包含扩增KRAS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。

原理

KRAS Pyro Kit用于实时定量检测人KRAS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。KRAS基因编码一种在EGFR信号通路中有重要作用的蛋白。KRAS基因的突变会影响该蛋白对下游通路的激活。所有癌症中将近30%发生KRAS基因突变，其中直肠癌（35%）和肺癌（18%）。

可检测以下种类的突变：

编码子12/13：G12A、G12C、G12D、G12R、G12S、G12V和G13D
编码子61：Q61E、Q61H、Q61L和Q61R

程序

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""和""）。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。推荐使用KRAS Plug-in Report进行分析。该报告确保使用正确的LODs和分析的不同序列可自动检测所有突变。也可在PyroMark Q24 System上使用其他分析工具进行分析（参见" Pyrogram trace of a GGT to GAT mutation in base 2 of codon 12"）。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 TGT mutation in base 1 of codon 12"和" Pyrogram trace of the reanalysis of the sample in the previous figure"）。

查看图表

Illustration of the KRAS assay.

Pyrogram trace of a normal genotype in codons 12 and 13.

Pyrogram trace of a GGT to GAT mutation in base 2 of codon 12.

Pyrogram trace of a GGT to TGT mutation in base 1 of codon 12.

Pyrogram trace of the reanalysis of the sample in the previous figure.

应用

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

可检测以下种类的突变：

编码子12/13：G12A、G12C、G12D、G12R、G12S、G12V和G13D
编码子61：Q61E、Q61H、Q61L和Q61R

辅助数据和图表

Pyrogram trace of a normal genotype in codons 12 and 13.

资源

For quantitative measurements of mutations in codons 12, 13, and 61 of the human KRAS gene

Version 1.3.0.6
For use with PyroMark Q24 Software version 2.0.8

Download Safety Data Sheets for QIAGEN product components.

For installation and use with PyroMark Q24 Instruments and PyroMark Q24 Software version 2.0

FAQ

The NRAS Pyro, EGFR Pyro. BRAF Pyro, KRAS 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

Mutation assays designed by QIAGEN detect the most common mutations. In addition, QIAGEN provides plug-ins to analyze more mutations and convenient reports for PyroMark KRAS, PyroMark EGFR, and PyroMark BRAF kits. The plug-ins can be obtained by emailing pyro.plugin@qiagen.com

If the mutation cannot be analyzed successfully, please send the original run files to QIAGEN Technical Service for further assistance.

FAQ ID -9065

Mutations in KRAS codon 61 must be analyzed separately with the KRAS Plug-in and not with the RAS extension plug-in.

FAQ ID - 3622

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

Paraffin-embedded tissue

Blood

QIAamp DNA Blood Mini Kit

FAQ ID -2380

Plug-Ins are available for:

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

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