Norgen Urine Exfoliated Cell and Bacteria RNA Purification Kit

Norgen Urine Exfoliated Cell and Bacteria RNA Purification Kit

SKU: 22550

For the rapid purification of total RNA (including microRNA) from exfoliated cells in urine.

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Features and Benefits

• Purify all sizes of RNA (including microRNA) without the need for phenol
• Isolate and detect total RNA from 1 mL up to 50 mL urine
• Provides high-quality RNA for sensitive applications — isolate RNA from as little as 100 cells
• Rapid processing time from sampling to downstream testing
• Purification is based on spin column chromatography using Norgen’s proprietary resin matrix

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This kit provides a rapid spin column method for isolating and purifying total RNA (including microRNA) from exfoliated cells shed into urine from the urinary tract. It purifies all sizes of RNA, from large mRNA and rRNA down to microRNA (miRNA) and siRNA. RNA is preferentially purified from proteins and contaminants (e.g., glucose, salts) without phenol or chloroform. The purified RNA is of high integrity and suitable for downstream applications such as real-time RT-PCR, qRT-PCR, Northern blotting, NGS, and expression arrays. Multiple samples can be processed in 20 minutes.

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Background

RNA biomarkers from exfoliated cells can be used as non-invasive tools for diagnostic and research applications, including detection and monitoring of bladder, kidney, or urinary-tract cancers.

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Supporting Data

Figure 1. Isolation and Detection of RNA from Exfoliated Cells in 50 mL of Urine
RNA was isolated from exfoliated cells in 50 mL urine from a healthy male using Norgen’s Urine (Exfoliated Cell) RNA Purification Kit. The bind, wash, and elute procedure was followed, and RNA was recovered in 20 µL Elution Buffer. HeLa cell dilution standards (1–1,000 cells/mL) were processed in parallel. Seven microliters of eluted RNA were reverse transcribed, and 1 µL of cDNA was amplified by real-time PCR targeting ribosomal S14.
The PCR baseline graph showed red lines (HeLa RNA standards), blue lines (RNA from 50 mL non-spiked urine), and yellow lines (no template).

OCR 그래프 설명:

• 그래프 제목: PCR Amplification vs Cycle - 23-Sep-06 15:43.opd
• X축: Cycle (10–36)
• Y축: PCR Base Line Subtracted CF RFU (0–4500)
• 범례: 🔺 Standards / 🔷 50 mL Plain Urine / 🟡 No template
• 주석: 1,000/mL, 100/mL, 10/mL, 1/mL
• 농도가 낮을수록 증폭 곡선이 오른쪽으로 이동함.

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Kit Specifications

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Storage Conditions and Product Stability

All solutions should be tightly sealed and stored at room temperature. The kit is stable for 2 years from shipment date.

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Documentation

• Protocol (22550) PDF
• Safety Data Sheet (SDS)
• Research Brochure

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FAQs

Poor RNA Recovery

1. Incomplete lysis — ensure proper Buffer SK and lysozyme addition.
2. Column clogging — do not exceed 50 mL urine or 1×10⁶ cells.
3. Use only supplied Elution Buffer E.
4. Ethanol must be added to lysate and Wash Solution A.
5. Low cell density or few bacteria may yield little RNA; increase input volume or use sensitive detection (BioAnalyzer, RT-PCR).

Column Clogging

• Caused by inefficient lysis, too many cells/bacteria, high DNA, or low centrifuge temperature (<20°C).
• Avoid exceeding input limits and ensure complete lysis.

RNA Degradation

• Prevent RNase contamination and work quickly.
• Use RNase-free lysozyme.
• Store RNA at –20°C (short-term) or –70°C (long-term).
• Use fresh urine samples.

Poor Downstream Performance

• Ensure double washing with Wash Solution A to remove salts.
• Perform dry spin to remove ethanol carryover.

Genomic DNA Contamination

• May result from high input material.
• Treat with RNase-free DNase I (e.g., Norgen RNase-Free DNase I Kit, Cat. 25710).

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Citations

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