Description
Formerly DNA Extraction Control 670.
qPCR Extraction Control not only enables users of a diagnostic qPCR assay to determine if there are inhibitors in the PCR assay, but also to validate the success of the extraction step, reducing the chance of obtaining a false negative result in the sample DNA.Product Highlights
- Simple - streamlined protocol for straightforward validation of DNA extraction and determination of qPCR assay inhibition
- Sensitive - control assay identifies even small effects on DNA extraction and inhibition of amplification
- Optimized - control DNA has a sequence with no known homology to any organism thereby avoiding detection of sample DNA
- Specific - probe-based assay designed specifically for real-time PCR assays
- Flexible - ideal for use with a wide range of sample types, including inhibitor-rich samples like blood, urine and sputum samples
Product Description
A common practice in qPCR is to add a known amount of spiked control DNA after DNA extraction, this monitors PCR inhibition but has no value as an extraction control. The ideal situation is to have the test sample and internal control undergo the same processing prior to qPCR. Meridian has developed the qPCR Extraction Control, which more closely mimics the test sample, as compared to spike controls. Genetic material from the test sample and the qPCR Extraction Control is simultaneously extracted by common extraction methods, with the extraction control being as sensitive to inhibition and extraction failure as the test sample.
The qPCR Extraction Control cells are of a known concentration, containing the Internal Control DNA sequence. This sequence contains no known homology to any organism and, importantly, has minimal interference with detection of sample DNA. The qPCR Extraction Control cells are spiked into the lysis buffer with the target sample, prior to DNA extraction. Control Mix, which includes primers and probe, is then added to the reaction mix before amplification. Signal derived from the Internal Control DNA confirms the success of the extraction step. qPCR Extraction Control also monitors co-purification of PCR inhibitors that may cause biased or false amplification patterns.
Applications
- Pathogen detection
- Cancer risk assessment
- Gene expression analysis
- Copy number variation (CNV) analysis
- Genotyping
- Viral loading

Application Note
qPCR Extraction Control Redebiomall.com
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多谢……
各位大侠:小弟最近做个长片段的CDNA与T载体的连接,片段大小6.4K,载体选用的是pUC19和pGEMT-easy,反转录酶用的是RevertAidTirstStrandcDNASynthesisKit(#K1621),CDNA第二链的扩增用的是LATaq。我是用纯病毒的RNA做模板,操作步骤严格按照说明做的,也曾经有一次获得过电泳图清晰显示的是全长6.4KB左右的片段,与载体连接,结果筛选到的几个克隆,插入的片段仅有2KB左右,奈何?可最近几次再用此反转录试剂盒却不能得到所需片段。在此,小弟想问问大家,你们是否遇到同样的问题,是如何解决的?所用的试剂(载体和酶的选择)和方法是什么,可以告诉后来者,共同进步吧。

