HiPure Universal RNA Kit

The HiPure Universal RNA Kit combines MagZol reagent lysis with silica membrane purification to enable efficient RNA extraction from difficult or heterogeneous tissue samples. The MagZol-based phenol–guanidine chemistry improves disruption of fibrous tissues, lipid-rich tissues and other challenging biological materials.

Following MagZol lysis, RNA is captured on a silica membrane column and purified through multiple washing steps to remove proteins, genomic DNA residues and organic contaminants. The workflow is suitable for tissue samples where conventional column-based RNA extraction may show reduced yield or purity.

For routine RNA purification workflows laboratories may use the HiPure Total RNA Plus Kit (R4111), while automated magnetic bead RNA purification is supported by the MagPure Universal RNA Kit (IVD3020).

Workflow

Workflow Overview

The HiPure Universal RNA Kit uses a MagZol lysis and phase-separation workflow followed by silica column purification for total RNA extraction from a broad range of biological samples. Samples are lysed in MagZol Reagent, a phenol / guanidine-based system designed for strong sample disruption and RNase inactivation. After chloroform addition and centrifugation, RNA is recovered in the upper aqueous phase, while much of the DNA, protein and organic-phase background is separated away. The aqueous RNA phase is then adjusted with Buffer RW2 and loaded onto a silica spin column, where RNA is captured, washed, dried and eluted in RNase-free water.

Sample Handling Logic

This workflow is suitable for animal tissue, plant material, fungal samples, bacteria, cultured cells and other sample types that benefit from stronger front-end lysis and phase-based cleanup before column purification. The main variation is concentrated in the sample disruption and MagZol homogenization stage. Tissue and plant samples may require mechanical homogenization or liquid-nitrogen grinding, while fungal, yeast or bacterial samples may require more intensive disruption. Once the aqueous phase has been recovered, the downstream silica column workflow remains consistent.

Time and Workflow Characteristics

Under typical manual operation, the workflow is usually completed within about 40–55 minutes, depending mainly on sample disruption quality, phase separation handling and repeated column loading volume. Compared with a direct column workflow, this route is more involved because of the MagZol and chloroform phase-separation step, but it provides stronger front-end cleanup for complex or difficult samples. For detailed step-by-step conditions, workflow guidance and estimated processing times, please refer to the Workflow Note in the Download section.

Specifications

Technical Validation

HiPure Universal RNA Kit was evaluated as a hybrid RNA extraction workflow combining MagZol-based phenol / guanidine lysis with silica membrane purification. The workflow uses MagZol Reagent for efficient sample disruption and RNase inactivation, followed by chloroform phase separation, ethanol-assisted RNA binding, column washing and elution. This design is intended to combine the broad sample compatibility of one-step extraction with the purity advantage of silica column cleanup.

In animal tissue testing, total RNA was extracted from 50 mg chicken tissue samples including liver, kidney, spleen, heart, muscle and lung. From 50 mg chicken liver input, RNA yields reached 169.2–181.5 µg, with A260/280 values of 2.07–2.11 and A260/230 values of 1.94–1.98. From 50 mg kidney and spleen inputs, RNA yields were 103.7–103.9 µg and 127.6–127.7 µg, respectively. Lower-yielding tissues such as muscle and lung produced 6.7–11.2 µg and 32.4–34.8 µg RNA under the tested conditions. Agarose gel electrophoresis showed clear RNA band patterns, supporting RNA integrity across the tested tissue types.

Plant sample compatibility was tested using 100 mg soybean, maize and paddy leaf inputs. From 100 mg soybean leaf input, the kit produced RNA yields of 72.34–81.02 µg with A260/280 values of 2.19–2.20. From 100 mg maize and paddy leaf inputs, RNA yields were 9.94–12.40 µg and 13.38–14.58 µg, respectively, with A260/280 values of approximately 2.06–2.12. Electrophoresis analysis showed visible large-molecular RNA bands, supporting the use of the MagZol-plus-column workflow across both animal and plant sample types.

A comparison with commonly used commercial RNA extraction workflows was performed using 50 mg liver and heart tissue inputs. From 50 mg liver input, R4130 produced RNA yields of 160.7–173.8 µg, with A260/280 values of 2.13–2.14 and A260/230 values of 2.12–2.16. The reference workflows produced liver RNA yields in a similar range, while R4130 showed higher A260/230 values under the tested conditions. From 50 mg heart input, R4130 produced RNA yields of 14.1–15.7 µg, with A260/280 values of 2.12–2.13 and A260/230 values of 1.34–1.52, supporting comparable recovery and acceptable purity for lower-yielding tissue samples.

Batch consistency was further evaluated using 10 mg pig liver input. The reference batch produced RNA yields of 61.18–62.15 µg, while the QC batch produced 61.61–62.72 µg. A260/280 values remained at 2.08–2.10 and A260/230 values were 2.03–2.25 across the tested batches. Electrophoresis analysis showed clear RNA bands without obvious degradation, supporting consistent extraction performance between the tested batches.

Application-oriented testing was performed using six different clinical tissue sample types, including heart valves, mesentery, leaf fat, liver, kidney and intestine. The R4130 workflow produced total RNA amounts of 0.73–42 µg, with RNA concentrations of 23–1203 ng/µL. RNA integrity assessment showed RIN values of 7.4–9.1 and 28S/18S ratios of 1.3–1.9 across the tested samples. These results support the use of R4130-prepared RNA as input for downstream applications requiring intact RNA, including sequencing-oriented library preparation workflows, while final library performance should still be interpreted together with sample condition and library QC results.

Kit Contents

Storage and Stability

MagZol Reagent 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 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.

Experiment Data

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.