Introduction
HiPure FFPE RNA Plus Kit extends the standard FFPE RNA purification workflow by integrating DNase digestion during RNA extraction. Removal of genomic DNA contamination improves reliability of downstream transcription and gene expression analysis.
The workflow maintains the same column-based RNA purification chemistry while incorporating a dedicated cleanup step to improve RNA purity.
For standard RNA purification workflows see HiPure FFPE RNA Kit - R4143.
Laboratories requiring simultaneous DNA and RNA purification from the same FFPE tissue digestion 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 (with DNase)
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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, DNase
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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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≥10μl
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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
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Efficient DNA removal – unique method to effectively remove genomic DNA
Kit Contents
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Contents
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R414402
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D414403
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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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DNase I
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600 µl
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5 x 600 µl
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DNase Booster Buffer
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1.5 ml
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6 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. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 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.
Experiment Data
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:
