Total RNA Purification Micro Kit – 35300

Total RNA Purification Micro Kit: High-Quality Total RNA from Very Low Inputs

The Total RNA Purification Micro Kit extracts total RNA from low cell number samples and elutes it in a concentrated 20 µL volume. It suits small cell inputs, needle biopsies, laser-captured microdissection (LCM) samples, circulating tumour cells (CTCs) and other precious material where every nanogram counts.

Purification uses spin column chromatography built on Norgen’s proprietary resin separation matrix. All RNA sizes, from large mRNA and lncRNA down to microRNA (miRNA), are recovered in a single fraction without a phenol step. The resulting RNA has excellent RIN values and A260/A280 ratios, so it is ready for qRT-PCR, RT-PCR, microarrays, NGS and other downstream work.

Key Benefits:

• Sensitivity: Works down to a single cell. The kit is sensitive and linear down to a few cells without the need for carrier RNA, so your downstream results reflect your sample alone.
• Complete RNA Recovery: All sizes in one fraction. RNA is bound and eluted irrespective of size, and the column binds small RNAs such as miRNA regardless of their GC content. There is no phenol step, so you don’t need to split fractions or run separate small RNA preparations.
• Concentrated Eluate: Elute in just 20 µL. A small elution volume keeps low-yield samples at a workable concentration for RT-qPCR and library preparation.
• Speed and Simplicity: Ten purifications in 20 minutes. The spin column format is quick and needs no specialist equipment beyond a microcentrifuge.
• Versatile Sample Compatibility: One kit for many specimens. It handles animal cells, animal tissues and LCM samples, and the purified RNA is suitable for small RNA sequencing.

Technical Insights

Figure 1. Great Isolation Sensitivity

The Total RNA Purification Micro Kit allows sensitive RNA extraction from as little as a single cell. Total RNA was extracted from a decreasing number of HeLa cells. Five µL of the eluted RNA was used as the template in a 20 µL RT-qPCR reaction to detect the human S14 transcript. The S14 transcript was detected from as little as a single cell.

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

The kit allows sensitive but high quality RNA extraction from small inputs such as LCM samples. Total RNA was isolated from six different LCM samples, and RNA yield and quality (260/280 ratio) were assessed by NanoVue spectrophotometry. The kit recovered good amounts of RNA with an excellent 260/280 ratio.

Figure 3. Better Efficiency of RNA Recovery at Single Cell Input

The kit allows sensitive RNA extraction from as little as a single cell and outperforms a competitor’s kit that claims similar sensitivity. Total RNA was extracted from a decreasing number of HeLa cells using the Total RNA Purification Micro 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 transcripts. RNA isolated with the Total RNA Purification Micro Kit showed much higher relative expression of each transcript tested at every cell input, down to a single cell.

Specifications

• Pack size: 50 preps (manufacturer catalogue number 35300)
• Technology: Spin column chromatography using Norgen’s proprietary resin separation matrix
• Size of RNA purified: All sizes, including small RNA (< 200 nt)
• Maximum column binding capacity: 35 µg
• Maximum column loading volume: 650 µL
• Minimum elution volume: 20 µL
• Maximum starting material: 5 x 10⁵ animal cells; 3 mg animal tissue (for most tissues, fibrous tissue requires an additional Proteinase K treatment); up to 5 x 10⁵ cells for LCM
• Average yield: ~1.5 µg from 1 x 10⁵ HeLa cells
• Time to complete 10 purifications: 20 minutes
• Kit contents: Buffer RL (40 mL), Wash Solution A (38 mL), Elution Solution A (6 mL), 50 Micro Spin Columns, 50 Collection Tubes, 50 Elution Tubes (1.7 mL) and a product insert
• Not supplied: 96 to 100% ethanol (90 mL must be added to Wash Solution A before first use)
• Storage and stability: Store all solutions tightly sealed at room temperature. Stable for 2 years from the date of shipment.
• For research use only. Not intended for in vitro diagnostics.