QIAamp Circulating Nucleic Acid Kit

从人类血浆或血清中浓缩和纯化游离DNA和RNA

  • 浓缩的核酸,高上样量,低洗脱体积
  • 高效回收DNA和RNA碎片
  • 无需有机提取或乙醇沉淀
  • 完全去除污染物和抑制剂

QIAamp Circulating Nucleic Acid Kit大大简化了从血浆或血清中浓缩和纯化游离DNA和RNA的过程。

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QIAamp Circulating Nucleic Acid Kit (50)
For 50 preps: QIAamp Mini Columns, Tube Extenders (20 ml), QIAGEN Proteinase K, Carrier RNA, Buffers, VacConnectors, and Collection Tubes (1.5 ml and 2 ml)
55114
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Carrier RNA(poly A) 12x1350µg-15-25°C,KG
Carrier RNA(poly A) 12x1350µg-15-25°C,KG
1017647
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The QIAamp Circulating Nucleic Acid Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Reproducible recovery of circulating DNA.|Improved recovery of fragmented DNA.|Efficient purification of circulating miRNA.|Efficient recovery of methylated DNA.|
DNA fragments (200 bp and 1000 bp) in equal amounts were added to 24 plasma samples. DNA was purified from 5 ml plasma using the QIAamp Circulating Nucleic Acid Kit, with an elution volume of 80 μl. DNA yield was quantified by duplex, real-time PCR of 66 bp targets specific for each DNA fragment using the QuantiTect Multiplex PCR Kit.|Circulating DNA was purified from 5 ml plasma using the QIAamp Circulating Nucleic Acid Kit or from 1 ml plasma using the QIAamp MinElute Virus Vacuum Kit, with elution volumes of 100 μl. DNA yield was quantified by duplex, real-time PCR of a 66 bp amplicon and a 500 bp amplicon within the 18S rRNA gene using the QuantTect Multiplex PCR Kit. The difference between amplification of the 500 bp amplicon and 66 bp amplicon shows that the DNA is fragmented, resulting in a lower abundance of intact 500 bp target sequences compared with the shorter 66 bp target. (Note that the difference is greater than the fivefold difference in sample input volumes.)|Circulating RNA was purified from 4 ml pooled plasma using the QIAamp Circulating Nucleic Acid Kit, with an elution volume of 50 μl. The standard kit protocol was compared with the specialized protocol for miRNA. miRNA 30b yield was quantified using a TaqMan microRNA assay (Applied Biosystems), and miRNA 16 yield was quantified using a Human miScript Assay (QIAGEN). The lower CT values with the miRNA protocol indicate higher yields of the miRNA species.|Spiked methlyated DNA and non-target poly-A RNA were purified from 4 ml plasma using the QIAamp Circulating Nucleic Acid Kit, with an elution volume of 45 μl. Bisulfite conversion was carried out using the EpiTect Bisulfite Kit (QIAGEN), and methylation-specific PCR (MSP) was performed using real-time MSP assays and components of the QuantiTect Multiplex PCR Kit on the ABI PRISM 7900HT Sequence Detection System. Data from two fully methylated loci were analyzed.|
Performance

分析血液中肿瘤特异性的胞外DNA片段和mRNA可对普通血液样品进行特异性地肿瘤类型检测。这些存在于血清或血浆中的游离核酸通常都是<1000 bp(DNA)或<1000 nt(RNA)的片段。此外,小至21 nt的miRNAs可作为某些癌症和疾病状态的生物标记。

QIAamp Circulating Nucleic Acid Kit可从人类血浆、血清和其他无细胞体液中高效纯化游离核酸。纯化效率高、重现性好、产量高(参见"Reproducible recovery of circulating DNA")。应用延长的上样管和真空装置QIAvac 24 Plus可将样品处理量提至5 ml,洗脱体积可在20–150 μl之间灵活变化,可浓缩低浓度核酸。 该试剂盒采用先进的技术,硅胶膜选择性的结合目标核酸,提高片段化核酸的回收率(参见"Improved recovery of fragmented DNA")。

使用RNase-Free DNase Set进行DNA柱上消化,纯化获得不含DNA的游离RNA 。特别设计的流程确保高效纯化诸如miRNA等小RNA(参见"Efficient purification of circulating miRNA")。使用QIAamp Circulating Nucleic Acid Kit高效纯化甲基化DNA,经纯化的DNA维持其甲基化状态,可进行亚硫酸氢盐转化实验,进行甲基化状态分析(参见"Efficient recovery of methylated DNA")。

经纯化和浓缩的游离DNA和RNA不含蛋白、核酸酶和其他杂质,可即用于各种下游应用,包括:

PCR 和定量real-time PCR以及RT-PCR
用于基于血液样品的癌症生物标记的研究、验证和检测
病毒核酸检测 
Principle

QIAamp Circulating Nucleic Acid Kit简化了从血浆或血清中纯化游离DNA和RNA的过程。无需苯酚-氯仿抽提。核酸特异性结合到QIAamp Mini离心柱上,污染物流走。PCR抑制物,如:二价阳离子和蛋白,可通过三步有效的洗涤步骤被完全去除,结合在离心柱上的纯核酸可用水或试剂盒中的缓冲液洗脱。使用QIAamp Circulating Nucleic Acid技术从人类血浆、血清或尿液中获得游离的DNA和RNA。

QIAamp Circulating Nucleic Acid Kit有可选择性结合的硅胶膜和20–150 μl灵活的洗脱体积。使用RNase-Free DNase Set进行DNA柱上消化,可纯化游离RNA。

Procedure

QIAamp Circulating Nucleic Acid的流程包括4步(裂解、结合、洗涤和洗脱),使用QIAamp Mini离心柱在真空装置上处理。简单的操作适用于同时处理多个样品,可在2小时内处理24个样品。QIAamp Circulating Nucleic Acid Kit有待使用者验证是否能适用于所有的病毒种类的RNA和DNA纯化。

Applications

QIAamp Circulating Nucleic Acid Kit可从含低浓度DNA和RNA碎片的原始样本(人血浆中常规游离DNA的浓度为1–100 ng/ml)中纯化和浓缩游离的DNA、RNA、miRNA,包括病毒核酸。起始样本体积至多5 ml。

QIAamp Circulating Nucleic Acid Kit可从以下样本类型中纯化和浓缩核酸:

人类血浆
血清
尿液
Feature
Specifications
Applications PCR, real-time PCR
CE/FDA/IVD compatible -
Elution volume 20–150 µl
Format QIAamp Mini Columns
Main sample type Serum, plasma
Processing Manual (vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Free-circulating DNA, RNA and miRNA, viral DNA, viral RNA
Sample amount 1-5 ml
Technology Silica technology
Time per run or per prep <2 hour for 24 preps
Yield Varies, due to donor-to-donor variations and disease status

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Kit Handbooks
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For concentration and purification of free-circulating DNA and RNA from human plasma or serum
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