Total RNA Purification Kit (96-Well Format)

In stock
SKU
1024300
681,00 €
Total RNA Purification Kit (96-Well Format)
Supplier
Norgen Biotek
Cat. no.
1024300
Kit size
2 plates (2x96 wells)
Links

Protocol
Information Sheet

Features

  • Isolate a diversity of RNA species - All RNA species can be isolated, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).
  • Isolate RNA from a wide range of samples - RNA can be isolated from cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi and plants (Figure 1).
  • No phenol or chloroform extractions - Total RNA, including all small RNA species, is isolated without the use of harmful chemicals such as phenol or chloroform.
  • Isolate RNA from very small samples - Total RNA has been isolated and detected from as little as a single animal cell (Figure 3).
  • Fast and easy processing - 96-Well plates can be rapidly processed in 30 minutes using either a vacuum manifold or centrifugation format.
  • Recovered RNA is suitable for many downstream applications - 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.

Norgen’s Total RNA Purification 96-Well Kit provides a rapid method for the high-throughput isolation and purification of total RNA from cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi and plants. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), without the use of phenol or chloroform.
Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. The RNA is preferentially purified from other cellular components such as proteins without the use of phenol or chloroform. The purification could be performed on either a vacuum manifold or using centrifugation. The process involves first lysing the cells or tissue of interest with the provided Lysis Solution (please see the flow chart on page 4). Ethanol is then added to the lysate, and the solution is loaded onto the 96-Well Filter Plate. Norgen’s resin binds RNA in a manner that depends on ionic concentrations. Thus only the RNA will bind to the resin in the wells, while the contaminating proteins will be removed in the flowthrough or retained on the top of the resin. The bound RNA is then washed with the provided Wash Solution in order to remove any remaining impurities, and the purified total RNA is eluted with the Elution Buffer. The purified RNA is of the highest integrity, and can be used in a number of downstream applications. * average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.
User-Friendly Procedure



Binding capacity per well 50 µg
Maximum loading volume per well 500 µl
Size of RNA purified All sizes including
Time to complete 96 purifications 30 minutes
Average RNA yield HeLa cells (1x10exp6 cells): 15 µg E.coli (1x10exp9 cells): 50 µg
Isolate Total RNA from a Diversity of Species
Total RNA was isolated from 8 mg of brain (Lanes A), kidney (Lanes B), liver (Lanes C), lung (Lanes D) and spleen tissue (Lanes E), 7.5 x 10^5 HeLA (Lanes F) and CHO cells (Lanes G), and 5 x 10^8 bacteria (Lanes H) using Norgen’s Total RNA Purification 96-Well Kit. From observing the formaldehyde-agarose gel, it can been seen that Norgen’s kit can be used to successfully isolate total RNA, including small RNA species, from a broad range of sample types.











Consistent Isolation of Total RNA, Including microRNA
Total RNA was isolated from samples of 5 x 10^5 HeLa cells using Norgen’s Total RNA Purification 96-Well Kit. Aliquots of each total RNA sample were then used in 2 different RT-qPCR reaction. In the first RT-qPCR the S15 gene of mRNA was amplified (Panel A), and in the second RT-qPCR reaction the miR-21 microRNA was amplified (Panel B). Both the mRNA and microRNA were amplified in a consistent manner from all the samples, with low variability of the C T values. Thus both types of RNA are being consistently isolated using this kit.

  • Quantitative, real-time PCR
  • RT-PCR
  • Northern blotting
  • RNase protection and primer extension
  • Expression array assays
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