筛查FFPE肺组织中肿瘤相关的PDGFRA1基因突变
A/使用MDX168提取缓冲液处理含有从肿瘤血管生成相关的PDGFRA1突变肺肿瘤FFPE组织 (突变型),然后取1.3 ng DNA直接进行qPCR检测; 或Qiagen QIAamp DNA FFPE组织提取试剂盒对等量的突变型FFPE组织进行DNA提取,然后分别使用B/ KAPA Probe Force qPCR Mix (Roche)和C/ TaqPath ProAmp Multiplex Master Mix (ThermoFisher)对1.3ng 提纯的DNA进行qPCR检测。纯合子等位基因G (红色),等位基因A (蓝色),杂合子等位基因G/A (绿色),NTC (黑色)和x表示未知。结果显示,与对提纯的DNA检测相比,Lyo-ReadyTM可冻干基因分型FFPE直扩qPCR预混液 (MDX168) 对FFPE组织粗提取物的检测具有更强的等位基因识别和更紧密的簇集。
Lyo-Ready Genotyping Direct qPCR FFPE, 4x, MDX168
更好的提取缓冲液,对粗糙的 FFPE 样品有更大的耐受性。与所有用于检测基因变异(如单核苷酸和拷贝数变异)的双标记探针化学试剂兼容,可提供高重复性的等位基因鉴别。
文档与资源
SNPs (single nucleotide polymorphisms) or point mutations are the most common types of genetic variation and comprise the major part of phenotype diversity between individuals. SNPs can be responsible for both resistance or susceptibility to a certain disease and once individual SNPs have been identified by sequencing, they can be used for pharmacogenetics, in evaluating and predicting a patient’s response to treatment and risk of adverse events, or for diagnostics, such as in cystic fibrosis screening. Although blood is an ideal source for genotyping analysis, formalin-fixed paraffin-embedded (FFPE) tumor tissues are also widely used and constitute valuable resources for retrospective studies due to the possibility of long-term storage at room temperature. Despite the extremely time-consuming and labour-intensive workflows, success rate of genotyping assays using DNA purified from such samples remains low.
To minimize the workflow to isolate DNA from FFPE tissue, Meridian has created the Lyo-Ready Genotyping Direct qPCR FFPE, this allows digestion of the tissue in a single tube, without the need for purification, greatly reducing the risk of sample damage, loss and contamination whilst simplifying and accelerating the analysis process. Furthermore, it can be used in a liquid format or lyophilized format to create ambient-temperature stable assays, making it ideal for high throughput.
产品资料
可冻干基因分型FFPE
直扩qPCR预混液可冻干基因分型FFPE
直扩qPCR预混液
Lyo-Ready Genotyping Direct qPCR BloodLyo-Ready Genotyping Direct qPCR Blood
常见问答: Lyo-Ready 可冻干基因分型血液直扩qPCR预混液
等位基因区分是在一个特定的基因位点上区分两个或多个序列变化的方法。通常,一个等位基因(“野生型”DNA序列)是常见的,而其他等位基因(突变)是罕见的。在一个SNP中有3个基因型:一个二倍体纯合子(两个野生型等位基因(PP)),一个二倍体纯合子(两个突变型等位基因(PP))和一个杂合子(一个野生型等位基因和一个突变型等位基因(PP))。
SNPs(单核苷酸多态性)有助于预测个体对某些药物的耐药性或易感性(药物遗传学),对毒素等环境因素的易感性,以及致病风险风险。SNPs也可以用来追踪家族内疾病相关基因变异的遗传。
qPCR (Real-time PCR) enables you to screen known SNPs. The benefits of qPCR are that it is easy, accurate, and can scale to high throughput. The bioinformatic analysis is also less complex than for other technologies, such as sequencing and microarrays.
5 ' -核酸酶(TaqMan)等位基因鉴别使用一对引物来扩增目标区域和两个等位基因特异性探针来检测目标SNP等位基因,从而可得知样本的基因型。与正常的qPCR不同,在基因分型中,扩增子的荧光值仅在PCR结束时测量,结果显示在散点图上,因此可以允许上百个检测同时进行检测。
FFPE样本受到多种因素的影响,比如说固定时导致DNA交联、组织固定的时间、年份长短和组织的储存条件。这这些因素导致从FFPE样本中提取DNA很耗时耗力,提取出的DNA容易出现碎片化,用来做基因分型检测的成功率很低。
Yes, the Lyo-Ready Genotyping Direct qPCR FFPE can be used as a liquid mix or lyophilized and stored at ambient temperature without needing to change the reaction conditions and lyophilization/storage will not affect the sensitivity of the test.
可以单独采购MDX004 Tissue Extract-PCR Buffers
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