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PAXgene Tissue DNA Kit

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

  • 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

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。

Cat No./ID: 767134
PAXgene Tissue DNA Kit (50)
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

仅限于科研应用,不适用于诊断,不为疾病的诊断、预防或治疗提供参考信息。


Specifications for fixation and storage conditions in PAXgene Tissue Fix and PAXgene Tissue Stabilizer were determined using animal tissues.



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Significant hands-on time savings compared to the manual purification.

Comparison of sample preparation times between manual and QIAcube procedures for 12 PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissues. Hands-on time without incubation times.

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从固定并稳定在PAXgene Tissue Containers中的不同类型组织中纯化高品质DNA。
来自不同器官的组织用PAXgene Tissue Fix固定3小时,转移到PAXgene Tissue Stabilizer中在室温下储存24小时。处理完成后,使用PAXgene Tissue DNA Kit从石蜡包埋的组织切片中纯化DNA。M:标记。
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Yield of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed (PF) tissue types, processed on the QIAcube.

Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissues, processed with the PAXgene Tissue DNA Kit on the QIAcube in one single run. DNA yield per 10 mg tissue.

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Absorbance ratio of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed, paraffin-embedded (PF) tissue types, processed on the QIAcube.

Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissue, processed with the PAXgene Tissue DNA Kit on the QIAcube in a single run. Ratio of absorbance at 260 and 280 nm.

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Comparison of manual and automated procedure: DNA yield from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.

DNA yield from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube or manually.

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Comparison of manual and automated procedure: Absorbance ratio from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.

Ratio of absorbance at 260 and 280 nm from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube or manually.

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High-quality DNA from PFPE tissue with preserved morphology.

[A] Hematoxylin and eosin (H&E) staining of PAXgene Tissue-fixed, paraffin-embedded (PFPE) human colorectal cancer tissue and [B] DNA on agarose gel electrophoresis using 0.8% TBE buffered gels with 200 ng genomic DNA isolated in triplicate from 5 cases (#1–5) of human PFPE colorectral cancer. M: markers.

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PAXgene Tissue DNA实验流程。
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DNA without chemical modification can be used for demanding downstream applications.

Multiplex and long-range PCR of DNA from PAXgene Tissue-fixed, paraffin-embedded (PFPE) human colorectal cancer tissue (modified according to Viertler et al., (2012) A new technology for stabilization of biomolecules in tissues for combined histological and molecular analyses. J. MOl. Diagn. 14, 458). [A] Multiplex PCR of 8 different genomic DNA fragments ranging from 22 to 955 bp. [B] Long-range PCR of a 5 kb genomic DNA fragment.

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Agarose gel electrophoresis of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed (PF) tissue types, processed on the QIAcube.

Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissue, processed with the PAXgene Tissue DNA Kit on the QIAcube in one single run. DNA from each tissue type (300 ng) on agarose gel electrophoresis using 1% TBE buffered gels; M: markers.

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Comparison of manual and automated procedure: Agarose gel electrophoresis from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.

Agarose gel electrophoresis from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube and manually. DNA from each replicate and tissue type (200 ng) on agarose gel electrophoresis using 1% TBE buffered gels; M: markers.

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")。

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
Features
Specifications
应用 PCR, qPCR, real-time PCR
洗脱体积 45–70 µl
规格 96-well plate
主要样本类型 Tissue samples
处理 Manual (centrifugation or vacuum)
样本量 25–100 mg
技术 Silica technology
Time per run Varies
产量 Varies

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产品介绍与指南 (3)
Two worlds in one sample
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For isolation and purification of genomic DNA from tissue samples stabilized using the PAXgene Tissue System 

 


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Moving toward excellence and standardization in tissue collection and fixation

 


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试剂盒操作手册 (1)
For isolation and purification of genomic DNA from tissue samples stabilized in PAXgene Tissue Containers
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参考文献
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