PAXgene Tissue DNA Kit

从固定和稳定于PAXgene Tissue Container的组织中纯化DNA

S_2516_PAXTissueDNA
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PAXgene Tissue DNA Kit (50)

Cat. No. / ID:  767134

For 50 DNA preps: PAXgene DNA Mini Spin Columns, Processing Tubes, Microcentrifuge Tubes, Carrier RNA, and Buffers; to be used in conjunction with PAXgene Tissue Containers
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使用动物组织确定在PAXgene Tissue Fix和PAXgene Tissue Stabilizer中固定和贮存条件规范。
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。
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Features

  • 固定、稳定和纯化的完整系统
  • 从组织中纯化高品质DNA
  • 保持组织形态

Product Details

固定和储存于PAXgene Tissue Container的组织样本可后续经石蜡包埋,用于病原体研究和后续的DNA、RNA和/或miRNA研究时,使用PAXgene Tissue DNA Kit从固定和稳定于PAXgene Tissue Container的组织中纯化DNA,纯化基于离心柱形式的硅胶膜DNA纯化技术。配合容器,该试剂盒为分子分析提供从采样、固定和稳定到高品质DNA纯化的完整解决方案。

Performance

应用PAXgene Tissue DNA Kit纯化的总DNA纯度非常高,DNA A260/A280比为1.7–1.9,同时吸光度扫描显示其在260 nm处有对称峰,证实是高纯度的基因组DNA。污染最小化,并且纯化所得DNA可直接用于下游应用,无检测到的PCR抑制。采集器和试剂盒配合使用,为分子分析提供从采样、固定和稳定到高品质DNA纯化的完整解决方案(参见"High-quality DNA from different tissue types fixed and stabilized in PAXgene Tissue Containers")。

Principle

目前传统组织学所使用的组织固定方法对分子分析的应用带来限制。含有福尔马林的固定剂与生物分子交联,修饰核酸和蛋白质。在组织固定、储存和处理过程中,交联导致核酸降解。因为无法完全去除交联,其引起的化学修饰将抑制敏感的下游应用,如定量PCR或RT-PCR。为了实现对同一样本进行分子病理检测和传统病理检测,需要一种方法在固定分子的同时保存组织形态。

PreAnalytiX研制了PAXgene Tissue System以满足这些需求。该系统包括组织采集装置(PAXgene Tissue Container用于采集、固定、储存及运输人类组织样本)以及用于纯化总RNA、DNA或miRNA的试剂盒。PAXgene Tissue Container可进行组织固定,用于组织病理学研究;也可从同一样本中纯化高纯度核酸,用于分子分析。固定和稳定方法保持了组织形态学,福尔马林固定组织中同时存在没有破坏性交联和降解的完整核酸。

为了分离基因组DNA,该体系需要应用PAXgene Tissue Containers采集和稳定组织,然后应用PAXgene Tissue DNA Kit分离和纯化DNA。

Procedure

PAXgene Tissue Container是预装有2种试剂的双腔容器。PAXgene Tissue Fix组织固定液快速浸润并固定组织。组织固定后,将其从PAXgene Tissue Fix中移出,转移到同一容器中含有PAXgene Tissue Stabilizer稳定液的另外一个腔内。组织储存在PAXgene Tissue Stabilizer稳定液中,其核酸和组织形态可在室温下稳定3–7天,在2–8℃下可稳定2–4周,不同组织的保存时间不同。储存于–15–30℃时,组织形态和核酸完整性可能稳定至少26个月。

固定了的样本可包埋于石蜡中,用于组织学研究。从PAXgene Tissue固定、石蜡包埋(PFPE)的组织样本中纯化核酸可在石蜡包埋之前或之后进行。

PAXgene Tissue DNA Kit提供3种不同操作流程,用于从PAXgene Tissue Containers固定和稳定的组织中提取基因组DNA。

组织裂解在裂解缓冲液、Buffer TD1中进行,使用蛋白酶K进行消化。缓冲液条件应结合Buffer TD2和乙醇进行调整,确保提供最佳的DNA结合条件,然后将裂解液装到PAXgene DNA离心柱上。在离心过程中,DNA会选择性地结合到硅胶膜上,污染物则流过柱子。分别用洗涤缓冲液TD3和TD4进行2次有效洗涤,去除剩余的污染物和酶抑制剂,然后用低盐洗脱缓冲液Buffer TD5洗脱DNA,可即用于下游应用(参见" The PAXgene Tissue DNA procedure")。

See figures

Applications

纯化所得DNA可即用于各种下游应用,包括:

PCR和定量real-time PCR
Southern印迹
药物基因组研究
SNP发现和SNP基因分型

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR, qPCR, real-time PCR
Format96-well plate
Main sample typeTissue samples
ProcessingManual (centrifugation or vacuum)
Time per runVaries
Elution volume45–70 µl
Sample amount25–100 mg
YieldVaries
TechnologySilica technology

Resources

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
Technical Information and Important Notes (2)
This note is to inform you that the street address for PreAnalytiX GmbH has changed from “Feldbachstrasse” to “Garstligweg 8”. Please be informed that the update of the product labeling to the new address is ongoing.
In order to reduce paper consumption and oblige the growing number of customers requesting an environmentally friendly alternative to traditionally printed handbooks, we are now providing kit handbooks for Research Use Only (RUO) PreAnalytiX kits on our website only.
产品介绍与指南 (3)
Moving toward excellence and standardization in tissue collection and fixation

 

Two worlds in one sample
For isolation and purification of genomic DNA from tissue samples stabilized using the PAXgene Tissue System 

 

试剂盒操作手册 (1)
Safety Data Sheets (1)
Certificates of Analysis (1)

Publications

Comprehensive DNA Methylation and Extensive Mutation Analyses of HER2-Positive Breast Cancer.
Yamaguchi T; Mukai H; Yamashita S; Fujii S; Ushijima T;
Oncology; 2015; 88 (6):377-84 2015 Jan 14 PMID:25591616
Next-gen tissue: preservation of molecular and morphological fidelity in prostate tissue.
Gillard M; Tom WR; Antic T; Paner GP; Lingen MW; VanderWeele DJ;
Am J Transl Res; 2015; 7 (7):1227-35 2015 Jul 15 PMID:26328007
Assessment of PAXgene Fixation on Preservation of Morphology and Nucleic Acids in Microdissected Retina Tissue.
Liu Y; Edward DP;
Curr Eye Res; 2016; 42 (1):104-110 2016 Jul 13 PMID:27409982
A Critical Evaluation of the PAXgene Tissue Fixation System:  Morphology, Immunohistochemistry, Molecular Biology, and Proteomics.
Mathieson W; Marcon N; Antunes L; Ashford DA; Betsou F; Frasquilho SG; Kofanova OA; McKay SC; Pericleous S; Smith C; Unger KM; Zeller C; Thomas GA;
Am J Clin Pathol; 2016; 146 (1):25-40 2016 Jul PMID:27402607
The focus on sample quality: Influence of colon tissue collection on reliability of qPCR data.
Korenkova V; Slyskova J; Novosadova V; Pizzamiglio S; Langerova L; Bjorkman J; Vycital O; Liska V; Levy M; Veskrna K; Vodicka P; Vodickova L; Kubista M; Verderio P;
Sci Rep; 2016; 6 :29023 2016 Jul 7 PMID:27383461

FAQ

Where can I find additional information for PreAnalytiX PAXgene products?
You can find additional information relating to the PreAnalytiX PAXgene products on the PreAnalytiX website .
FAQ ID - 3515
Is a special processing protocol needed?

To prevent biomolecule degradation during processing, dehydration must begin with at least 70–100% ethanol. We recommend using low-melting paraffin (melting point ≤56°C) and do not incubate samples in liquid paraffin for more than 3 hours.

Processing protocols for the PAXgene Tissue System are listed in the appendices of the PAXgene Tissue Container Product Circular and PAXgene Tissue FIX Container (50 ml) Product Circular.
FAQ ID -2523
What is the RT-PCR performance of RNA purified from PAXgene Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissue compared to RNA from snap frozen or formalin-fixed, paraffin-embedded (FFPE) tissue?
The PAXgene Tissue fixation and stabilization chemistry is free of cross-linking reagents. RNA purified from PFPE and PFCE is free of chemical modifications and performs similarly or identically to RNA isolated from frozen tissue. For examples of the correlation of gene expression levels in snap frozen tissue, FFPE, and PFPE, see Figure 4 in Groelz et al., Exp Mol Pathol. 2013 Feb; 94(1) and Figure 3 in Viertler et al., J Mol Diagn. 2012 Sep; 14(5).
FAQ ID -2538
What is the purity of nucleic acids extracted with the PAXgene Tissue kits?

The PAXgene Tissue DNA, RNA, and miRNA Kits are based on proven QIAGEN technologies. Nucleic acids isolated with these kits are generally of high purity.

On average, measurements of the A260/A280 ratio for DNA purified with the PAXgene Tissue DNA Kit are >1.7, and ratios for RNA and miRNA purified with the PAXgene Tissue RNA or miRNA Kits are both >1.8.

For examples of RNA purity see Technical Note "Yield, purity, and integrity of RNA purified from PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue" under the Product Resources tab.

FAQ ID -2533
Is it possible to microdissect PAXgene Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissues?
Yes. Supplementary protocols for generating sections from PAXgene Tissue fixed, Paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissue blocks for manual and laser microdissection are available under the Product Resources tab.
FAQ ID -2531