For purification of microRNA and total RNA from formalin-fixed, paraffin-embedded tissue sections
- Effective purification of miRNA and total RNA
- Novel method to overcome formalin crosslinking
- Efficient release of RNA without compromising integrity
- Streamlined protocol providing RNA in just 85 minutes
The miRNeasy FFPE Kit enables purification of total RNA that includes RNA from approximately 18 nucleotides upwards from formalin-fixed, paraffin-embedded (FFPE) tissue sections. The kit provides recovery of usable RNA fragments, including miRNA and other small RNAs, for applications such as quantitative, real-time RT-PCR. Procedures can be automated on the QIAcube Connect.
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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
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217504
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miRNeasy FFPE Kit适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。
从FFPE组织中高效纯化RNA。|卓越的产量和实验表现。|高效的real-time RT-PCR。|miRNeasy FFPE实验流程。|
使用miRNeasy FFPE Kit或酚-氯仿抽提从图示的大鼠组织中纯化包括miRNA在内的总RNA。使用miScript PCR System和纯化的RNA作为针对miRNA miR-16的定量real-time RT-PCR分析的模板。结果显示,对于肺和肝脏组织,使用miRNeasy FFPE Kit纯化后的CT值更低,说明其miRNA纯化产量高于使用酚-氯仿抽提的产量。对于肾脏组织,使用两种方法的CT值相当。对于所有检测组织类型,使用miRNeasy FFPE Kit纯化后的CT值均更低或相当。|使用miRNeasy FFPE Kit或Supplier A的同类试剂盒,从图示的大鼠组织的FFPE切片中纯化RNA。[A] 测量260 nm处的吸光值,确定RNA的产量。[B] 使用miScript PCR System和纯化的RNA作为针对miRNA miR-16的定量real-time RT-PCR分析的模板。结果显示,使用miRNeasy FFPE Kit纯化,CT值更低,说明miRNA的产量高于使用Supplier A的试剂盒纯化的产量。|采用福尔马林固定大鼠肝脏组织24小时或60小时,然后使用miRNeasy FFPE Kit纯化包括miRNA在内的总RNA。使用miScript PCR System和纯化的RNA作为模板进行定量real-time RT-PCR分析,检测miRNA miR-16和miR-29a以及PGK1基因的较大mRNA。结果显示,从同一洗脱液中可同时检测到上述两种miRNA和mRNA。||
原理
Fixing tissues with formalin leads to RNA-RNA and RNA-protein crosslinking which impairs RNA performance in downstream applications. The miRNeasy FFPE Kit provides special lysis and incubation conditions to reverse formalin crosslinking of RNA. A specially developed lysis buffer efficiently releases RNA from tissue sections while avoiding further RNA degradation. The lysate is treated with DNase then optimized binding conditions allow purification of all usable RNA from approximately 18 nucleotides upwards.
操作流程
The entire miRNeasy FFPE procedure can be completed in as little as 85 minutes. The optimized lysis buffer allows sample lysis with proteinase K digestion in only 15 minutes without sacrificing RNA yields. After lysis, samples are incubated at 80ºC for 15 minutes to reverse formalin crosslinking. Genomic DNA is then rapidly removed in an optimized DNase step with DNase Booster Buffer, and concentrated RNA is purified using RNeasy MinElute spin columns (see flowchart " miRNeasy FFPE procedure"). Since RNA is eluted in a volume of just 14–30 µl, smaller reaction volumes are possible in downstream applications.
应用
The miRNeasy FFPE Kit allows purification of miRNA with total RNA for use in a variety of applications, such as quantitative, real-time RT-PCR.
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特点
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参数
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应用
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PCR, qPRC, real-time RT-PCR, microArray
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洗脱体积
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14-30 µl
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规格
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Spin column
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主要样本类型
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FFPE tissue samples
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处理
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Manual (protocol for automated processing on QIAcube available)
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纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白
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miRNA, total RNA
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样本量
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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)
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技术
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Silica technology
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产量
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Varies
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Critical factors for molecular analysis of FFPE samples
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Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow
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浏览
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High-quality, nucleic acid purification for successful PCR and NGS experiments.
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FAQ ID -1737
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浏览
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FAQ ID -2801
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浏览
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I received one of the kits in the list below containing the MinElute columns, however they were left out for a while and not stored at 2-8°C upon receipt. Can I still use them? AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro
FAQ ID - 3560
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浏览
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FAQ ID -828
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浏览
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FAQ ID -2797
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浏览
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FAQ ID -2798
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浏览
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FAQ ID -3002
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浏览
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For purification of total RNA, including miRNA, from formalin-fixed, paraffin-embedded tissue sections
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显示更多
|
图片
从FFPE组织中高效纯化RNA。
使用miRNeasy FFPE Kit或酚-氯仿抽提从图示的大鼠组织中纯化包括miRNA在内的总RNA。使用miScript PCR System和纯化的RNA作为针对miRNA miR-16的定量real-time RT-PCR分析的模板。结果显示,对于肺和肝脏组织,使用miRNeasy FFPE Kit纯化后的CT值更低,说明其miRNA纯化产量高于使用酚-氯仿抽提的产量。对于肾脏组织,使用两种方法的CT值相当。对于所有检测组织类型,使用miRNeasy FFPE Kit纯化后的CT值均更低或相当。
卓越的产量和实验表现。
使用miRNeasy FFPE Kit或Supplier A的同类试剂盒,从图示的大鼠组织的FFPE切片中纯化RNA。[A] 测量260 nm处的吸光值,确定RNA的产量。[B] 使用miScript PCR System和纯化的RNA作为针对miRNA miR-16的定量real-time RT-PCR分析的模板。结果显示,使用miRNeasy FFPE Kit纯化,CT值更低,说明miRNA的产量高于使用Supplier A的试剂盒纯化的产量。
高效的real-time RT-PCR。
采用福尔马林固定大鼠肝脏组织24小时或60小时,然后使用miRNeasy FFPE Kit纯化包括miRNA在内的总RNA。使用miScript PCR System和纯化的RNA作为模板进行定量real-time RT-PCR分析,检测miRNA miR-16和miR-29a以及PGK1基因的较大mRNA。结果显示,从同一洗脱液中可同时检测到上述两种miRNA和mRNA。
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