Single Cell RNA Purification Kit – 51800

Single Cell RNA Purification Kit: High-Sensitivity Isolation for Critical Samples

The Single Cell RNA Purification Kit is a specialised tool engineered for the challenging isolation of total RNA from extremely low-input biological samples. Whether working with single cells, laser-capture microdissection (LCM) samples, or small populations of sorted cells, this kit ensures the recovery of high-integrity RNA without the losses associated with standard precipitation methods. It is the ideal choice for researchers requiring a representative snapshot of the transcriptome from scarce material.

The system utilises a proprietary resin-based spin column chromatography technology that binds RNA efficiently irrespective of size or concentration. This allows for the simultaneous isolation of all RNA species—from large mRNA and ribosomal RNA down to microRNA (<200 nt) and siRNA. By eliminating the need for phenol/chloroform extraction and employing a specially designed micro spin-column, the protocol concentrates high-purity RNA into elution volumes as low as 8 µl, ensuring the concentration is sufficient for sensitive downstream applications like qPCR and Next-Generation Sequencing (NGS).

Key Benefits:

• Broad Dynamic Range: Scalable from 1 to 200,000 cells. Maintains linear recovery across a wide range of inputs, from a single cell to 2 x 10⁵ cells or LCM samples.
• Maximum Concentration: 8 µl elution volume. The micro spin-column design minimises dead volume, allowing for highly concentrated elution ready for direct use in low-volume reactions.
• Comprehensive Capture: Total RNA including miRNA. Unlike silica kits that lose small molecules, this resin captures all RNA sizes, providing a complete picture of gene regulation.
• Rapid Workflow: 20 minutes for 10 samples. A streamlined “bind-wash-elute” process reduces bench time and limits the exposure of samples to potential RNase contamination.
• Phenol-Free Chemistry: Safe and clean isolation. Avoids toxic organic solvents, resulting in pure RNA free from inhibitors that could compromise downstream enzymatic reactions.

Technical Insights

Figure 1. Better Efficiency of RNA Recovery at Single Cell Input

Norgen’s Single Cell RNA Purification Kit allows sensitive RNA extraction from as little as a single cell and outperforms competitor’s kits that claim similar sensitivity. Total RNA was extracted from a decreasing number of HeLa cells using Norgen’s Single Cell RNA Purification Kit and a competitor’s spin column purification kit. The extracted RNA was subjected to RT-qPCR to detect the human S15 and 5S rRNA transcript in the isolated RNA. RNA isolated by Norgen’s Single Cell RNA Purification Kit showed much higher relative expression of each RNA transcript tested at all cell input numbers down to single cell.

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Figure 2. High Quality RNA Isolated from Laser-Captured Microdissection (LCM)

Norgen’s Single Cell RNA Purification Kit allows sensitive but high-quality RNA extraction from small inputs such as Laser-Captured Microdissection (LCM). Total RNA was isolated from 6 different LCM samples. RNA yield and quality (260/280 ratio) were assessed by NanoVue spectrophotometry. Norgen’s Single Cell RNA Purification Kit recovered good amounts of RNA with an excellent 260/280 ratio.

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Figure 3. Consistent and Linear Recovery of All Sizes of RNA down to Single Cell Input

Norgen’s Single Cell RNA Purification Kit allows sensitive RNA extraction from as little as a single cell with great linearity. Total RNA was extracted from a decreasing number of HeLa cells using Norgen’s Single Cell RNA Purification Kit. The extracted RNA was subjected to RT-qPCR to detect all sizes of RNA, including the human S15 (mRNA), 5S rRNA transcript (small RNA), and miR-21 (microRNA). RNA isolated by Norgen’s Single Cell RNA Purification Kit showed very high consistency (over 99%) of recovery of each RNA transcript tested at all cell input numbers down to a single cell.

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Specifications

• Starting Material: Single Cell to 2 x 10⁵ cultured cells; Laser-Captured Microdissection (LCM) samples.
• Column Binding Capacity: 10 µg.
• Minimum Elution Volume: 8 µl.
• Processing Time: ~20 minutes for 10 samples.
• Size of RNA Purified: All sizes, including small RNA (< 200 nt).
• Average Yield (HeLa): ~1.5 µg (from 1 x 10⁵ cells); >1 pg (from 1 cell).
• Shelf Life: Stable for 2 years at room temperature.