从福尔马林固定、石蜡包埋的组织切片中进行miRNA和总RNA的纯化

  • 有效纯化miRNA和总RNA
  • 创新方法用于克服福尔马林造成的交联
  • 有效释放RNA而不会破坏其完整性
  • 85分钟流畅的操作得到RNA

miRNeasy FFPE Kit确保从福尔马林固定、石蜡包埋(FFPE)的组织切片中纯化含约18个核苷酸以上的总RNA。试剂盒能够回收可用的RNA片段,包括miRNA和其他小RNA,用于定量real-time RT-PCR等下游应用。

Product Product no. Cat. no. List price:
 
 
Show details
    varies
Can't order online?
To place an order via phone, email or for requesting a quote, please provide the product’s name, number and catalog number.
Product Cat. no. List price:
miRNeasy FFPE Kit (50)
50 RNeasy MinElute Spin Columns, Collection Tubes, Proteinase K, RNase-Free DNase I, DNase Booster Buffer, RNase-Free Buffers, RNase-Free Water
217504
Inquire
The miRNeasy FFPE Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
  • Main Image Navi
Efficient RNA purification from FFPE tissues.|Superior yields and performance.|Effective real-time RT-PCR quantification.|miRNeasy FFPE procedure.|
Total RNA including miRNA was purified from the indicated rat tissues using either the miRNeasy FFPE Kit or phenol-chloroform extraction. Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-16 using the miScript PCR System. Results showed that for lung and liver tissues, CT values were lower after purification using the miRNeasy FFPE Kit, indicating that higher amounts of miRNA were purified than when using phenol-chloroform extraction. For kidney tissue, CT values were similar for both methods. CT values were similar or lower after purification using the miRNeasy FFPE Kit for all tissue types tested.|RNA was purified from FFPE sections of the indicated rat tissues using either the miRNeasy FFPE Kit or a similar kit from Supplier A. [A] RNA yields were determined by measuring absorbance at 260 nm. [B] Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-16 using the miScript PCR System. Results showed that CT values were lower after purification using the miRNeasy FFPE Kit, indicating that higher amounts of miRNA were purified than when using the kit from Supplier A.|Rat liver tissue was formalin fixed for 24 hours or 60 hours, followed by purification of total RNA including miRNA using the miRNeasy FFPE Kit. Purified RNA was used as a template in quantitative, real-time RT-PCR using the miScript PCR System to detect miRNAs miR-16 and miR-29a and for the larger mRNA of the PGK1 gene. Results show successful detection of both miRNAs as well as for the mRNA from the same eluate.||
Performance
使用miRNeasy FFPE Kit纯化的总RNA和miRNA的产量和性能优于采用其他方法纯化的miRNA,如使用苯酚-氯仿抽提或者其他供应商的类似试剂盒(参见"Effective real-time RT-PCR quantification"、"Efficient RNA purification from FFPE tissues"和"Superior yields and performance")。
Principle

用福尔马林固定的组织导致RNA-RNA和RNA-蛋白质交联,从而影响了RNA在下游应用中的表现。miRNeasy FFPE Kit提供了独特的裂解和孵育条件,可逆转福尔马林对RNA造成的交联。特别研制的的裂解缓冲液有效地将RNA从组织切片中释放出来,同时避免了RNA的进一步降解。裂解产物用DNase消化,然后优化的结合条件能够纯化含约18个以上核苷酸的所有可用的RNA。

Procedure
所有的miRNeasy FFPE操作流程都可在85分钟之内完成。优化的裂解缓冲液能够用蛋白质酶K消化样本裂解产物,仅需15分钟而且不损耗RNA产量。裂解后,在80°C下孵育样本15分钟逆转福尔马林交联。然后,在优化的DNase消化步骤中,用DNase Booster Buffer快速去除基因组DNA,并且用RNeasy MinElute离心柱纯化浓缩的RNA(参见"miRNeasy FFPE procedure")。由于RNA的洗脱体积仅有14–30 µl,在下游应用中可实现较小的反应体积。
Applications

miRNeasy FFPE Kit能够纯化包含miRNA的总RNA,并且适用于多种下游应用,如定量real-time RT-PCR。

Feature
Specifications
Applications PCR, qPRC, real-time RT-PCR, microArray
Elution volume 14-30 µl
Format Spin column
Main sample type FFPE tissue samples
Processing Manual (protocol for automated processing on QIAcube available)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein miRNA, total RNA
Sample amount up to 4 sections, each with a thickness up to 10 µm and a surface area up to 250mm^2 (automated protocol on QIAcube: up to 2 sections)
Technology Silica technology
Yield Varies

You are not authorized to download the resource

Kit Handbooks
1
For purification of total RNA, including miRNA, from formalin-fixed, paraffin-embedded tissue sections
Show details