HiPure Total RNA Plus Kit

The HiPure Total RNA Plus Kit (R4111) is designed for purification of high-quality total RNA from animal tissues and cultured cells using silica membrane spin column technology. The system integrates an additional genomic DNA removal step that eliminates genomic DNA contamination prior to RNA purification, enabling reliable RNA preparation for downstream gene expression analysis.

The extraction workflow combines efficient sample lysis with silica membrane RNA adsorption to selectively purify RNA molecules while removing proteins, genomic DNA and other contaminants. The resulting RNA is suitable for RT-PCR, transcriptomic analysis and other RNA-based molecular biology workflows.

Within the Magen RNA extraction systems, the HiPure Total RNA Plus Kit serves as a core column-based solution for high-purity RNA preparation when removal of genomic DNA contamination is critical for downstream applications.

Workflow

Workflow Overview

The HiPure Total RNA Plus Kit uses a dual-column silica workflow for purification of RNA from cultured cells, animal tissues and plant samples. Following disruption and chemical lysis, the lysate is first passed through a gDNA removal column so that genomic DNA is reduced before RNA binding is established. The RNA-containing flow-through is then adjusted with ethanol and loaded onto the RNA column, where RNA is captured, washed, dried and eluted in RNase-free water.

Sample Handling Logic

This workflow is designed for routine tissue and cell RNA extraction where operational simplicity and upstream gDNA removal are important. The main sample-dependent variation occurs during disruption and lysis, especially for tissue and plant material. Once the lysate has been cleared and passed through the DNA removal column, the downstream RNA-binding, washing and elution steps follow a consistent column workflow.

Time and Workflow Characteristics

Under typical manual operation, the workflow is usually completed within about 25–45 minutes, depending mainly on sample disruption and lysate viscosity. This route is suitable for laboratories that require a straightforward dual-column RNA workflow with gDNA reduction before RNA purification. For detailed step-by-step conditions, workflow guidance and estimated processing times, please refer to the Workflow Note in the Download section.

Specifications

Engineering Characteristics

Integrated genomic DNA removal

The workflow includes a dedicated gDNA removal column that eliminates genomic DNA contamination before RNA purification, reducing the need for additional DNase treatment.

Efficient RNA binding chemistry

Silica membrane adsorption enables selective binding of RNA molecules while cellular proteins and other contaminants are removed during washing steps.

Optimized lysis and inhibitor removal

The extraction buffers efficiently disrupt cellular structures and remove contaminants that may interfere with downstream reverse transcription reactions.

Spin-column workflow

The purification procedure is compatible with routine laboratory centrifuges and supports rapid RNA purification workflows in research laboratories.

Technical Validation

HiPure Total RNA Plus Kit was evaluated as a rapid silica column-based total RNA extraction workflow with integrated genomic DNA removal. The kit uses a DNA Mini Column before RNA binding, enabling total RNA purification from cultured cells and animal tissues without phenol / chloroform extraction or isopropanol precipitation. The purified RNA is intended for downstream applications such as RT-PCR, Northern blotting, poly(A)⁺ RNA purification, nuclease protection and in vitro translation.

In protein- and lipid-rich tissue testing, RNA was extracted from 10 mg chicken liver and 10 mg chicken brain samples. From 10 mg chicken liver input, the kit produced RNA yields of 45.46–48.24 µg, with A260/280 values of 2.11–2.12 and A260/230 values of 1.75–2.23. From 10 mg chicken brain input, RNA yields were 4.29–4.55 µg, with A260/280 values of 2.13–2.18 and A260/230 values of 1.90–2.15 under the tested conditions.

A comparison with a reference column-based RNA extraction workflow was performed using chicken liver input amounts of 5 mg, 10 mg, 20 mg and 40 mg. With 40 mg chicken liver input, R4111 produced RNA yields of 147–156 µg, compared with 147 µg from the reference workflow. With 20 mg input, R4111 produced 95–98 µg RNA, compared with 84 µg from the reference workflow. With 10 mg input, R4111 produced 41–47 µg RNA, compared with 37 µg from the reference workflow, while 5 mg input produced 19–21 µg RNA under the tested conditions.

Across the tested chicken liver input range, R4111 maintained A260/280 values of approximately 2.11–2.15 and A260/230 values of approximately 2.07–2.28. Agarose gel electrophoresis showed intact RNA band patterns and no obvious genomic DNA contamination, supporting the effectiveness of the gDNA removal column for routine DNA reduction before RNA purification.

The kit was also evaluated in a rapid cultured-cell extraction workflow using 5 × 10⁶ cultured cells per sample. Six cell samples were processed in approximately 30 minutes, and electrophoresis analysis showed clear total RNA bands with good integrity. These results support R4111 as a fast total RNA extraction workflow for routine animal tissue and cultured-cell samples where reduced genomic DNA background and short hands-on processing time are required.

For a broader view of Magen tissue and cell RNA extraction routes, the following resources may help place this product within the complete workflow system.