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).

Specifications

Principle

Tissue samples (10-100mg) are homogenized in MagZol Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the spin column, where the total RNA (up to 100µg) binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 30-100µl of RNase-free water.

Advantages

• Efficient DNA removal - one step RNA extraction can effectively remove genomic DNA
• High quality - one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast - several samples can be extracted in 30 minutes
• High applicability - samples including animals, plants, bacteria, cells, etc.
• High output - enable to process large samples and the yield can be up to 200μg

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

Choosing the appropriate RNA extraction kits depends on sample type, RNA species and workflow requirements.

For detailed guidance across tissue and cell RNA extraction workflows, refer to the Tissue/Cell RNA Kits Selection Guide.