HiPure Total RNA Kit

The HiPure Total RNA Kit provides a single-column silica membrane workflow for purification of large RNA molecules (>200 nt) from cultured cells and soft tissues. The method relies on chaotropic lysis followed by selective RNA adsorption on a silica membrane and sequential washing steps to remove proteins, salts and other contaminants.

The streamlined workflow is widely used for routine RNA preparation in molecular biology laboratories and supports downstream applications such as RT-PCR, qPCR and gene expression analysis. The protocol is suitable for standard laboratory workflows requiring consistent RNA purification from cultured cells or soft animal tissues.

Within the Magen RNA extraction systems, laboratories requiring genomic DNA removal during RNA purification may refer to the HiPure Total RNA Plus Kit (R4111), while automated RNA purification workflows can be implemented using the MagPure Universal RNA Kit (IVD3020).

Specifications

Principle

HiPure RNA technology simplifies total RNA isolation. Samples are first lysed and then homogenized. Ethanol is added to the lysate to provide ideal binding conditions. The lysate is then loaded onto the HiPure silica membrane and RNA binds to the silica membrane, and all contaminants are efficiently washed away. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using a convenient on-column DNase treatment. Pure, concentrated RNA is eluted in water.

Advantages

• Efficient removal of DNA - unique genomic DNA removal column without DNase treatment
• High quality - high purity total RNA can be directly used in various sensitive downstream applications
• Fast - several samples can be extracted in 20 minutes by column method
• Safe - no phenol chloroform extraction required
• Sensitive - RNA can be recovered at the level of PG

Technical Validation

HiPure Total RNA Kit was evaluated using animal and plant tissue samples to assess RNA yield, purity and integrity in a silica membrane spin-column workflow. The validation included manual extraction from 20 mg animal tissues and 100 mg plant tissues, with purified RNA analyzed by Nanodrop and agarose gel electrophoresis.

In animal tissue testing, RNA was extracted from kidney, spleen and liver samples and compared with a reference column-based RNA extraction workflow. HiPure Total RNA Kit produced RNA yields of 56–60 µg from kidney, 50–52 µg from spleen and 100–105 µg from liver under the tested conditions. The measured A260/280 values were 2.06–2.11, and A260/230 values were generally within 2.06–2.23, supporting recovery of RNA with suitable purity for routine downstream analysis.

Plant tissue extraction was further evaluated using pepper leaves, pumpkin leaves and Pachira macrocarpa leaves. From 100 mg plant input, the kit produced RNA yields of 55.47–70.31 µg from pepper leaves, 76.26–77.20 µg from pumpkin leaves and 46.13–50.31 µg from Pachira macrocarpa leaves. A260/280 values were 1.97–1.99, with most A260/230 values above 2.2 in the tested R4011 samples.

Electrophoresis analysis showed intact RNA band patterns from both animal and plant samples, supporting RNA integrity after extraction. The kit format also supports optional on-column DNase treatment for applications sensitive to residual genomic DNA, and the purified RNA is suitable for downstream workflows such as RT-PCR, Northern blotting, poly(A)⁺ RNA purification, nuclease protection and in vitro translation.

Kit Contents

Storage and Stability

The Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

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.