Norgen Cytoplasmic & Nuclear RNA Purification Kit

Norgen Cytoplasmic & Nuclear RNA Purification Kit

SKU: 21000
For 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 free of DNA and ready for direct use in RT-PCR/qRT-PCR
• Purification based on spin column chromatography using Norgen’s proprietary resin matrix

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 sizes of RNA, including small RNAs, without phenol. The kit includes reagents for 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations. Ten samples can be processed in approximately 45 minutes. Also available in a 100 prep size.

Background

Fractionated RNA isolation is useful when studying mature cytoplasmic RNA or nuclear pre-processed RNA. The cytoplasmic fraction is free of genomic DNA, making it ideal for downstream applications requiring DNA-free RNA.

Supporting Data

Figure 1. Effective Separation of HeLa Cell Cytoplasmic & Nuclear RNA
Norgen’s kit effectively separates cytoplasmic and nuclear RNA from 0.75 million HeLa cells.
Panel A: High-quality cytoplasmic and nuclear RNA purified using Norgen’s kit.
Panel B: Genomic DNA visible only in nuclear fractions, absent in cytoplasmic RNA.
Optional on-column DNase treatment removes residual genomic DNA.

OCR 텍스트 (Figure 1)
Panel A (RNA Gel):
Nuclear (1–3) Cytoplasmic (4–6)
Panel B (DNA Gel):
Nuclear (1–3) Cytoplasmic (4–6)

Figure 2. Superior Separation Compared to Competitor
Norgen’s kit provides better separation of cytoplasmic and nuclear RNA from 0.8 million HeLa cells compared to a competitor’s product.
Panel A: Higher yields and integrity of RNA with Norgen’s kit.
Panel B: Genomic DNA contamination observed only in nuclear fraction with Norgen’s kit, but also in cytoplasmic fraction with competitor’s kit.

OCR 텍스트 (Figure 2)
| 구분 | Norgen | Competitor A |
|------|---------|--------------|
| A (RNA Gel) | C / N | C / N |
| B (DNA Gel) | C / N | C / N |

Figure 3. Genomic DNA-free Cytoplasmic RNA
RT-PCR using human beta actin primers confirms absence of genomic DNA contamination in cytoplasmic RNA.
Lane M: Marker; Lanes 1–3: negative control; Lanes 2–4: RT-PCR product (166 bp).

OCR 텍스트 (Figure 3)
M 1 2 3 4

Figure 4. Specific Separation and Amplification of Cytoplasmic and Nuclear Transcripts
RT-PCR using U2 snRNA and S14 primers demonstrates successful separation of cytoplasmic and nuclear RNA fractions.

OCR 텍스트 (Figure 4)
Panel A: Cytoplasmic → Nuclear (U2 snRNA)
Panel B: Cytoplasmic ← Nuclear (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
• SDS (50 Prep)
• SDS (100 Prep)

FAQs

Why do I have poor RNA recovery?
Possible causes include insufficient cell solubilization, column clogging, use of alternative elution solution, missing ethanol addition, or improper PBS washing.

Why is the column clogged?
May result from excessive starting material, poor solubilization, high genomic DNA content, or low centrifuge temperature.

Why is RNA degraded?
RNase contamination, slow processing, improper storage, or thawing of frozen samples before isolation.

Why is RNA not performing well in downstream applications?
Insufficient washing or ethanol carryover may interfere with downstream assays.

How to prevent genomic DNA contamination in cytoplasmic fraction?
Ensure the nuclear pellet is not disturbed when transferring supernatant.

Markers for cytoplasmic/nuclear differentiation (RT-PCR):
U2 snRNA (nuclear), S14 RNA (cytoplasmic).

Related Products

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

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