Introduction
The HiPure Universal miRNA Kit (R4310) is designed for enrichment and purification of small RNA molecules including miRNA from tissues, cultured cells and other biological samples. The extraction workflow combines MagZol reagent chemistry with silica membrane purification to efficiently recover small RNA fractions.
Unlike conventional RNA purification methods that primarily recover large RNA molecules, the HiPure Universal miRNA Kit enables selective enrichment of small RNA molecules while also allowing purification of larger RNA fractions when required.
The purified RNA fractions are suitable for downstream applications including miRNA expression analysis and small RNA sequencing.
Details
Specifications
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Features
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Specifications
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Main Functions
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Isolation miRNA and other small RNA molecules(18nt), from cultured cells and various animal and human tissues, using MagZol reagent and column
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Applications
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RT-PCR, Northern Blot, 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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Animal tissues, adherent cells, suspension cells, bacteria, etc
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Sample amount
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Eukaryotic culture cells: ≤ 10^7, Animal tissue:<100mg, Yeast culture cells:<5 x10^7, Bacteria:<10^9
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Elution volume
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≥15μl
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Time per run
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≤40 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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Extraction Principle
The kit combines phenol/guanidine-based lysis with silica membrane purification to enable efficient isolation of total RNA and enrichment of small RNA species including miRNA. During the lysis step, MagZol reagent disrupts cells and tissues while denaturing proteins and RNases, preserving RNA integrity. Subsequent chloroform extraction separates RNA into the aqueous phase while removing proteins, lipids, and most genomic DNA.
RNA purification is then achieved using silica membrane adsorption under ethanol-controlled binding conditions. When a higher ethanol concentration is applied, both large and small RNA molecules bind to the membrane, enabling recovery of total RNA including miRNA. Under lower ethanol conditions, larger RNA molecules preferentially bind to the first column while small RNAs remain in the flow-through. After increasing ethanol concentration, these small RNA species bind to a second column, allowing isolation of an miRNA-enriched fraction while preserving the larger RNA fraction for parallel purification.
Engineering Characteristics
Phenol/Guanidine RNA Stabilization Chemistry
MagZol reagent rapidly lyses cells and tissues while denaturing RNases, preserving RNA integrity and preventing degradation of small RNA molecules during extraction.
Ethanol-Controlled RNA Size Selection
Differential ethanol concentrations regulate RNA adsorption to silica membranes, enabling selective enrichment of small RNA fractions while larger RNA molecules are captured separately.
Efficient Recovery of Regulatory Small RNAs
The extraction chemistry and binding conditions are optimized to efficiently recover miRNA and other regulatory small RNA species that are frequently lost during conventional RNA purification workflows.
Technical Validation
Validation experiments using animal tissue samples demonstrated efficient recovery of small RNA fractions with purity ratios A260/280 around 1.9–2.1, indicating RNA of sufficient purity for downstream molecular biology applications.
Extraction experiments showed that small RNA molecules could be efficiently enriched while larger RNA fractions remained available for parallel purification workflows.
Electrophoresis analysis confirmed enrichment of small RNA fragments predominantly below ~50 nt, consistent with efficient recovery of miRNA molecules.
Small RNA yields of up to ~26–29 µg from 50 mg liver tissue were observed in validation experiments, demonstrating efficient recovery from biological samples.
Performance Comparison
Compared with conventional RNA purification systems designed primarily for large RNA (>200 nt), the Universal miRNA Kit provides improved recovery of small RNA molecules through ethanol-controlled binding and sequential silica purification.
Key advantages include:
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Efficient enrichment of small RNA fractions including miRNA
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Simultaneous recovery of large RNA (>200 nt) for parallel analysis
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Reduced loss of regulatory RNA molecules during purification
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Comparable RNA purity and yield to widely used commercial extraction systems
These features make the system suitable for applications requiring reliable recovery of regulatory RNA species.
Typical Applications
• miRNA expression analysis
• Small RNA sequencing
• Gene regulation studies
• RNA interference research
• Regulatory RNA biology
Related Products
• HiPure Total RNA Plus Kit (R4111) – purification of high-purity total RNA from tissues
• HiPure Universal RNA Kit (R4130) – purification of RNA including small RNA fractions
• MagPure Serum miRNA Kit (R6628) – purification of miRNA from plasma and serum samples
• MagPure Universal RNA Kit (IVD3020) – automated RNA purification from tissue and cell samples
Kit Contents
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Contents
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R431002
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R431003
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Purification Times
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50 Preps
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250 Preps
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HiPure RNA Mini Columns
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100
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2 x 250
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2ml Collection Tubes
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100
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2 x 250
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MagZol Reagent
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60 ml
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270 ml
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Buffer RWC
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20 ml
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80 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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RNase Free Water
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10 ml
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30 ml
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Storage and Stability
MagZol Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its 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.
