Introduction
RNA extraction from formalin-fixed paraffin-embedded tissues requires digestion conditions capable of releasing fragmented RNA molecules while minimizing degradation during purification.
HiPure FFPE RNA Kit applies silica membrane column technology to recover total RNA from FFPE tissue sections commonly used in oncology research laboratories.
The workflow integrates optimized digestion and column adsorption chemistry to support RNA purification suitable for downstream molecular analysis.
For RNA workflows requiring genomic DNA removal see HiPure FFPE RNA Plus Kit - R4144.
Laboratories performing combined DNA and RNA analysis from limited FFPE material may refer to MagPure FFPE DNA/RNA Kit - R6327.
Details
Workflow
Workflow Overview
The HiPure FFPE RNA workflow uses a silica column–based route for purification of RNA from FFPE sections. Following deparaffinization, proteinase K digestion and controlled heat treatment, RNA binding conditions are established and RNA is purified through membrane binding, washing, drying and elution. An optional DNase treatment route is included when reduced genomic DNA carryover is required for downstream RNA analysis.
Sample Handling Logic
This workflow is designed for FFPE RNA recovery, where controlled heat exposure and residual DNA management are especially important. FFPE RNA is often fragmented before extraction begins, so the workflow balances tissue digestion and heat-assisted release with conditions intended to avoid unnecessary additional stress. When downstream assays are sensitive to genomic DNA contribution, the DNase treatment route provides an additional control point.
Time and Workflow Characteristics
Under typical manual operation, the overall workflow usually requires about 1.5–2 hours, depending on whether DNase treatment is included and on sample handling conditions. For detailed step-by-step conditions, workflow guidance and estimated processing times, please refer to the Workflow Note in the Download section.
Specifications
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Features
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Specifications
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Main Functions
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Isolation total RNA from FFPE tissue and section samples
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Applications
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RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation
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Purification method
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Mini spin column
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Purification technology
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Silica technology
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Process method
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Manual (centrifugation or vacuum)
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Sample type
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FFPE tissue sample
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Sample amount
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6mg
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Yield
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20μg
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Elution volume
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≥20μl
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Time per run
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≤60 minutes
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Liquid carrying volume per column
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800µl
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Binding yield of column
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100µg
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Principle
This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer.
Advantages
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High quality - high purity total RNA can be directly used in various sensitive downstream applications
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Fast - several samples can be extracted in 60 minutes by column method
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Safe - no phenol chloroform extraction required
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Sensitive - RNA can be recovered at the level of PG
Kit Contents
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Contents
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R414302
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D414303
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Purification Times
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50 Preps
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250 Preps
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HiPure RNA Micro Columns
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50
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250
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2ml Collection Tubes
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50
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250
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Buffer DPS
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60 ml
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250 ml
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Buffer FRL
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15 ml
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60 ml
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Buffer RLC
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15 ml
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60 ml
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Buffer RWC*
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10 ml
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50 ml
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Buffer RW2*
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20 ml
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2 x 50 ml
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Protease Dissolve Buffer
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1.8 ml
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10 ml
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Proteinase K
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24 mg
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120 mg
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RNase Free Water
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10 ml
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20 ml
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Storage and Stability
Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
Purchase Guide
For guidance on selecting the most appropriate FFPE nucleic acid extraction system based on target analyte, workflow format and downstream application requirements:
👉 FFPE Nucleic Acid Extraction Purchase Guide
For a broader technical overview of FFPE DNA, RNA and DNA/RNA co-extraction workflow routes, processing logic and application-oriented route design:
👉 FFPE Nucleic Acid Extraction Workflows Explained
For detailed workflow structure, estimated processing time and route-specific handling logic across representative FFPE workflows:
