IDH1/2 RGQ PCR Kit

用于检测FFPE脑部肿瘤组织DNA中12种IDH1IDH2突变

S_1084_5_GEN_V2

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IDH1/2 RGQ PCR Kit (20)

Cat. No. / ID:  873001

For 20 reactions: 9 Primer and Probe Mixes, WT Control, Positive Control, Master Mix, Nuclease-Free Water
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仅作研究用途。不用于诊断程序。任何表述或表示都并非旨在提供诊断、预防或治疗疾病的信息。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 利用PCR发夹技术准确检测7种IDH1和5种IDH2突变
  • 利用ARMS PCR技术灵敏地鉴别3种突变
  • 可用于即用型解决方案的标准化分析
  • 可用于Rotor-Gene Q实时荧光定量PCR分析仪的简单的工作流程

Product Details

在Rotor-Gene Q 5plex HRM Instrument上采用IDH1/2 RGQ PCR Kit进行real-time PCR,可实现对12种IDH1IDH2基因突变的检测。试剂盒提供了所有必需的试剂,可快速且灵敏地实现IDH1IDH2突变的检测。突变检测引物和探针混合物将引物与探针相结合,同时扩增突变型和野生型目标序列,此为还有一个核苷酸,3'端添加了磷酸基团,以防止延伸(PCR发夹技术),它可以特异性地结合野生型目标序列。

Principle

利用IDH1/2 RGQ PCR Kit中的试剂可完成9个独立的扩增反应,检测12种突变:

  • IDH1基因的132和100密码子及IDH2基因的172密码子的3次总扩增反应
  • IDH1基因的132和100密码子及IDH2基因的172密码子的3次突变扩增反应
  • IDH1 R132H、IDH1 R132C和IDH2 R172K突变的3次突变特异性扩增反应
使用IDH1/2 RGQ PCR Kit检测IDH1IDH2突变
基因 突变 碱基变化 Cosmic ID



IDH1
Arg132His 395G>A COSM28746
Arg132Cys 394C>T COSM28747
Arg132Ser 394C>A COSM28748
Arg132Gly 394 C>G COSM28749
Arg132Leu 395G>T COSM28750
Arg132Val 394_395 CG>GT COSM28751
Arg100Gln 299G>A COSM88208


IDH2
Arg172Lys 515G>A COSM33733
Arg172Met 515G>T COSM33732
Arg172Trp 514A>T COSM34039
Arg172Ser 516G>T COSM34090
Arg172Gly 514A>G COSM33731
总反应混合物

Total Primers and Probe Mixes(PPM-Total)使用引物和探针扩增突变型和野生型目标序列。

突变检测反应混合物

突变检测引物和探针混合物将引物与探针相结合,同时扩增突变型和野生型目标序列,此为还有一个核苷酸,3'端添加了磷酸基团,以防止延伸(PCR发夹技术),它可以特异性地结合野生型目标序列。

当PCR模板包含野生型序列时,因具有更高的亲合力,3'-磷酸寡核苷酸相比PCR引物的结合更具优势。DNA聚合酶几乎无延伸,且几乎未观察到扩增。

当存在突变序列时,PCR引物结合相比3'-磷酸寡核苷酸结合更具优势,扩增将继续。

突变鉴定反应混合物

ARMS(Amplification Refractory Mutation System)利用DNA聚合酶鉴别PCR引物的3'端是否匹配,实现等位基因特异性的扩增。

当PCR引物完全匹配时,扩增效率最高。当3'碱基不匹配时,仅出现低水平的背景扩增。

Procedure

IDH1/2 RGQ PCR Kit提供了12种IDH1IDH2基因突变检测所需的全部试剂。使用九种引物和探针混合物同时扩增突变型和野生型目标序列。只需将模板DNA及适当的引物和探针混合物加入即用型PCR预混液中,在Rotor-Gene Q 5plex HRM Instrument上采用试剂盒手册所述的优化循环程序开始反应。对于DNA纯化,我们推荐使用QIAamp DNA FFPE Tissue Kit,参照IDH1/2 RGQ PCR Kit手册中给定的改进实验方案。

Applications

利用IDH1/2 RGQ PCR Kit可以从福尔马林固定、石蜡包埋(FFPE)的脑部肿瘤组织样本DNA中检测12种IDH1IDH2基因突变。

Resources

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (2)
For detection of 12 IDH1 and IDH2 mutations
For research use only. Not for use in diagnostic procedures
For use with Rotor-Gene Q 5plex HRM instrument
Safety Data Sheets (1)
Certificates of Analysis (1)
Kit Handbooks (2)
For detection of 12 IDH1 and IDH2 mutations
For research use only. Not for use in diagnostic procedures
For use with Rotor-Gene Q 5plex HRM instrument

FAQ

Can I use stained slide materials with IDH 1/2 RGQ PCR kit?

No.  The pathologist should only proceed with unstained material.

 

 

FAQ ID -3101
Can I perform the IDH 1/2 RGQ PCR test with slides containing less than 40% tumor cells?

No. To detect all mutations, it is required to have at least 40% tumor cells.

FAQ ID -3098
Can I use the QIAGEN deparaffinization solution with the IDH 1/2 RGQ PCR kit?
No. The QIAGEN deparaffinization solution does not work with our kit.
FAQ ID -3100