21000 - Cytoplasmic & Nuclear RNA Purification Kit

Features High-quality RNA. No DNAse required (cytoplasmic fraction). No phenol extractions.

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Ordering Information

Item Cat. No. Kit Size Price (USD) Cytoplasmic & Nuclear RNA Purification Kit 21000 50 $250.00

Product Description
Norgen’s Cytoplasmic & Nuclear RNA Purification Kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. In certain circumstances it is desirable to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic RNA for some studies on expression profiling, or nuclear RNA in order to study pre-processed RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA.

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Norgen’s resin binds RNA in a manner that depends on ionic concentrations. The cytoplasmic RNA is preferentially purified from the nuclear RNA and the other cellular components such as genomic DNA and proteins, without the use of phenol or chloroform. The purified RNA is of the highest quality and can be used in a number of downstream applications.

Kit Specifications
Column Binding Capacity 50 ug Maximum Column Loading Volume 600 uL Maximum Amount of Starting Material:    Animal Cells 3 x 10E6 cells    Animal Tissues 15 mg	Time to Complete 10 Purifications 45 minutes Size of RNA Purified All sizes Average Yield:     HeLa Cells (1 x 10E6 cells)- cytoplasmic 15 ug   HeLa Cells (1 x 10E6 cells) - nuclear ≤3.5 ug

Key Figures

Figure 1. HeLa Cell Cytoplasmic & Nuclear RNA High Quality Cytoplasmic & Nuclear RNA Purified Using Norgen's Kit. Note the integrity and quality of both cytoplasmic and nuclear RNA. Ten microliters of each elution were run on a 1.5% formaldehyde-agarose gel. Lane M is Norgen's RNA Ladder, lanes 1-3 contain nuclear RNA and lanes 4-6 contain cytoplasmic RNA.

Figure 2. Genomic DNA-free Cytoplasmic RNA RT-PCR was performed using human beta actin primers on cytoplasmic RNA isolated fom HeLa cells using Norgen's Cytoplasmic & Nuclear RNA Purification Kit. Lane M is the DNA ladder, lanes 1 and 3 are the negative control, lanes 2 and 4 show the expected 166 bp RT-PCR product. The lack of a product in lanes 1 and 3 indicate that no genomic DNA contamination is present in the isolated cytoplasmic RNA.

Figure 3. No Genomic DNA Contamination in Cytoplasmic Fraction Cytoplasmic and nuclear RNA isolated from HeLa cells using Norgen's kit was run on a 0.9% agarose gel. Genomic DNA is clearly visible in the nuclear RNA fractions (lanes 1-3), however, no genomic DNA can be detected in the cytoplasmic RNA fractions (lanes 4-6).

Figure 4. User-Friendly Procedure

Benefits

• Rapid spin-column format allows for the processing of 10 samples in 45 minutes.
• Purified cytoplasmic RNA can be used directly in RT-PCR reactions with no DNAse treatment required (Fig 2).
• Cytoplasmic and nuclear RNA is isolated without the use of harmful chemicals such as phenol or chloroform.
• All nuclear and cytoplasmic RNA species can be isolated, from large mRNA species down to microRNA (miRNA) and small interfering RNA (siRNA).
• Purified RNA can be used in a number of downstream applications including real-time PCR, reverse transcription PCR, Northern blotting, RNAse protection and primer extension, and expression array analysis requiring the use of intact RNA.

US: 650-320-8669