Norgen Cytoplasmic and Nuclear RNA Purification Kit

Norgen Cytoplasmic and Nuclear RNA Purification Kit

SKU: 37400
Convenient purification of cytoplasmic and nuclear RNA from cultured cells and tissues.

Features and Benefits

• Excellent separation and purification of cytoplasmic and nuclear RNA
• Convenient and fast spin column format
• High quality and yield of RNA
• Isolate full diversity of RNA (including microRNA) without phenol
• Purified RNA ready for any application (RT-PCR, qRT-PCR, RNA-Seq, arrays, etc.)
• Cytoplasmic RNA is free of DNA and ready for direct use in RT-PCR/qRT-PCR
• Based on spin column chromatography using Norgen’s proprietary resin separation matrix

Description

This kit provides a rapid method for isolating and purifying both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. It isolates all RNA sizes, including small RNA species, without phenol.
Contains reagents for 50 cytoplasmic RNA preps or 25 cytoplasmic + 25 nuclear RNA preps. Ten samples can be processed in approximately 45 minutes. Also available in a 100 prep size.

Background

Fractionated RNA isolation allows targeted studies on mature cytoplasmic RNA or nuclear pre-processed RNA. The cytoplasmic fraction is free from genomic DNA contamination, enabling precise downstream analyses.

Supporting Data

Figure 1. Effective Separation of HeLa Cell Cytoplasmic & Nuclear RNA
Norgen’s kit effectively separates cytoplasmic and nuclear RNA from HeLa cells.

• Panel A: High-quality RNA integrity shown on 1.5% formaldehyde-agarose gel.
• Panel B: DNA gel confirms genomic DNA only in nuclear fraction.
  Optional on-column DNase treatment removes nuclear DNA.

이미지 OCR 데이터 요약:
| Panel | Fraction | Sample No. | Gel Type |
|--------|-----------|-------------|-----------|
| A | Nuclear | 1, 2, 3 | RNA Gel |
| A | Cytoplasmic | 4, 5, 6 | RNA Gel |
| B | Nuclear | 1, 2, 3 | DNA Gel |
| B | Cytoplasmic | 4, 5, 6 | DNA Gel |

Figure 2. Superior Separation Compared to Competitor
Norgen’s kit provides better separation and higher RNA yield than competitor’s.

• Panel A: RNA gel comparison between Norgen and Competitor A (C: Cytoplasmic, N: Nuclear).
• Panel B: DNA gel shows genomic DNA only in nuclear fraction for Norgen kit.

이미지 OCR 데이터 요약:
| Panel | Supplier | Fraction | Gel Type |
|--------|-----------|-----------|-----------|
| A | Norgen | C, N | RNA Gel |
| A | Competitor A | C, N | RNA Gel |
| B | Norgen | C, N | DNA Gel |
| B | Competitor A | C, N | DNA Gel |

Figure 3. Genomic DNA-free Cytoplasmic RNA
RT-PCR performed on cytoplasmic RNA shows expected product with no genomic DNA contamination.
Lane markers: M (Marker), 1–4 (Sample lanes).

Figure 4. Specific Separation and Amplification of Cytoplasmic and Nuclear Transcripts
RT-PCR using human U2 snRNA (nuclear) and S14 (cytoplasmic) primers confirms effective separation.

이미지 OCR 데이터 요약:
| Panel | Cytoplasmic | Nuclear | Marker |
|--------|--------------|----------|---------|
| A | (band) | (band) | U2 snRNA |
| B | (band) | (band) | S14 |

Kit Specifications

Storage Conditions and Product Stability

All solutions should be tightly sealed and stored at room temperature.
Stable for 2 years after shipment.

Kit Components

Documentation

• Protocol (50 prep)
• Protocol (100 prep)
• Application Note
• Safety Data Sheet (50 prep)
• Safety Data Sheet (100 prep)
• Poster

FAQs

Why do I have poor RNA recovery?

Possible causes include insufficient cell solubilization, column clogging, alternative elution buffer use, missing ethanol addition, or unwashed cell monolayer. Use recommended buffers and follow protocol precisely.

Why is the RNA degraded?

Potential causes: RNase contamination, slow procedure, improper RNA storage, thawed frozen samples, or improper tissue freezing. Maintain RNA integrity by working quickly and keeping samples cold.

Why is RNA not performing well in downstream applications?

Ensure three washes with Wash Solution A and perform dry spin to remove ethanol carryover.

How to prevent genomic DNA contamination in cytoplasmic fraction?

Avoid transferring nuclear pellet residues when separating cytoplasmic fraction.

Suggested RT-PCR markers

Refer to Figure 4: U2 snRNA (nuclear marker), S14 RNA (cytoplasmic marker).

Related Products

• Leukocyte RNA Purification Kit
• microRNA Purification Kit
• RNase-Free DNase I Kit
• Total RNA Purification Kit

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