HiPure Total RNA Kit

The HiPure Total RNA Plus Kit is designed for RNA purification workflows requiring stringent removal of genomic DNA contamination. The system integrates a DNA removal column with DNase digestion to ensure efficient elimination of residual genomic DNA during RNA purification.

The workflow supports purification of total RNA including small RNA species from tissue samples and cultured cells. The protocol is commonly used in RNA-seq experiments, RT-PCR analysis and other applications where trace genomic DNA contamination may affect downstream results.

For routine RNA extraction workflows laboratories may refer to the HiPure Total RNA Plus Kit (R4111), while automated RNA purification systems can be implemented using the MagPure Universal RNA Kit (IVD3020).

Workflow

Workflow Overview

The HiPure Total RNA Kit with DNase I uses a dual-column workflow with additional DNase-supported RNA purification. After disruption and lysis, genomic DNA is first reduced by passing the lysate through a gDNA removal column. The RNA-containing flow-through can then be directed into either a larger RNA route or a total RNA route including miRNA. Following RNA binding on the silica column, on-column DNase digestion provides an additional level of DNA background control before washing, drying and elution.

Sample Handling Logic

This workflow is designed for samples where RNA recovery and DNA background control are both important. Soft tissues, fiber-rich tissues, cultured cells and blood-derived cell pellets may require different front-end handling before the shared downstream column workflow. The route selection is determined by the analytical target: the larger RNA route is used when recovery of RNA above approximately 200 nt is sufficient, while the total RNA route including miRNA applies stronger binding preparation to retain small RNA species as well.

Time and Workflow Characteristics

Under typical manual operation, the workflow is usually completed within about 45–75 minutes, depending on sample pretreatment, route selection and the DNase digestion step. The workflow is slightly longer than a routine dual-column RNA protocol, but provides more structured DNA background control and broader route flexibility. For detailed step-by-step conditions, workflow guidance and estimated processing times, please refer to the Workflow Note in the Download section.

Specifications

Kit Contents

Storage and Stability

Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 weeks) at room temperature (15–25°C) does not affect their 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.

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.