QIAamp Circulating Nucleic Acid Kit – cfDNA/cfRNA Isolation

从人类血浆或血清中浓缩和纯化游离DNA和RNA

S_1583_RPA_QA_s

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QIAamp Circulating Nucleic Acid Kit (50)

Cat. No. / ID:   55114

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)
KitAccessories
QIAamp Circulating Nucleic Acid Kit
Carrier RNA
QIAamp Circulating Nucleic Acid 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。

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的(再)订购

特点

  • 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

产品详情

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。

绩效

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 
查看图表

原理

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。

程序

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。

应用

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

辅助数据和图表

Specifications

FeaturesSpecifications
ApplicationsPCR, real-time PCR
CE/FDA/IVD compatible-
TechnologySilica technology
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinFree-circulating DNA, RNA and miRNA, viral DNA, viral RNA
Time per run or per prep<2 hour for 24 preps
FormatQIAamp Mini Columns
Main sample typeSerum, plasma
ProcessingManual (vacuum)
Sample amount1-5 ml
Elution volume20–150 µl
YieldVaries, due to donor-to-donor variations and disease status

资源

产品介绍与指南 (1)
试剂盒操作手册 (1)
For concentration and purification of free-circulating DNA and RNA from human plasma or serum
安全数据表 (2)
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.
应用说明 (1)
Digital PCR (dPCR) is a powerful technique that detects and quantifies ultra-rare mutations in a high background of wild-type cfDNA down to 0.1% variant allele frequency. Here, we describe end-to-end manual and automated workflows that enable accurate detection and absolute quantification of ultra-rare PIK3CA variants in cfDNA using the QIAcuity Digital PCR System.
Safety Data Sheets (1)
Certificates of Analysis (1)

Publications

Noninvasive whole-genome sequencing of a human fetus.
Kitzman JO; Snyder MW; Ventura M; Lewis AP; Qiu R; Simmons LE; Gammill HS; Rubens CE; Santillan DA; Murray JC; Tabor HK; Bamshad MJ; Eichler EE; Shendure J;
Sci Transl Med; 2012; 4 (137):137ra76 2012 Jun 6 PMID:22674554
Non-invasive prenatal measurement of the fetal genome.
Fan HC; Gu W; Wang J; Blumenfeld YJ; El-Sayed YY; Quake SR;
Nature; 2012; 487 (7407):320-4 2012 Jul 19 PMID:22763444
Frequent methylation and oncogenic role of microRNA-34b/c in small-cell lung cancer.
Tanaka N; Toyooka S; Soh J; Kubo T; Yamamoto H; Maki Y; Muraoka T; Shien K; Furukawa M; Ueno T; Asano H; Tsukuda K; Aoe K; Miyoshi S;
Lung Cancer; 2011; 76 (1):32-8 2011 Nov 1 PMID:22047961
Implementing prenatal diagnosis based on cell-free fetal DNA: accurate identification of factors affecting fetal DNA yield.
Barrett AN; Zimmermann BG; Wang D; Holloway A; Chitty LS;
PLoS One; 2011; 6 (10):e25202 2011 Oct 4 PMID:21998643
IgH gene rearrangements as plasma biomarkers in Non- Hodgkin's lymphoma patients.
He J; Wu J; Jiao Y; Wagner-Johnston N; Ambinder RF; Diaz LA Jr; Kinzler KW; Vogelstein B; Papadopoulos N;
Oncotarget; 2011; 2 (3):178-85 2011 Mar PMID:21399237

FAQ

What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728
What is the average amount of DNA and RNA present in 1 ml normal serum?

According to an Interview with Professor Dennis Lo published in QIAGEN News Molecular Diagnostics, Issue No. 5, 2002, healthy individuals have about 500-1000 genome equivalents (DNA) per ml serum/plasma.

For free-circulating DNA in plasma, the concentration can range from 1–100 ng/ml in healthy individuals.

FAQ ID -635