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Total RNA Purification Kit (96-well format) for plants and fungi Add to Cart

Cat#: 31900-NB
Quantity: 2 plates/192 preps
Price: 839 €
Supplier: Norgen
Shipping: RT
User Manual  

For the rapid purification of total RNA (including microRNA) from plants and fungi

• Isolate RNA from a wide range of plant and filamentous fungi samples
• Isolate total RNA, including virus & viroid RNA – Purified RNA can be used for the detection of viral pathogens including viroids.
• Robust lysis buffer is well-suited to even challenging samples such as pine needle or grape leaf
• Isolate a diversity of RNA species - All sizes of RNA are isolated from large mRNA down to microRNA, without the use of phenol or chloroform
• Isolated RNA is of high quality, integrity and diversity
• Also available as 50 and 100 preps kit

Norgen’s Plant/Fungi Total RNA Purification Kit provides a rapid method for the isolation and purification of total RNA, including virus & viroid RNA, from a wide range of plants. Total RNA can be purified from fresh or frozen plant tissues, plant cells or filamentous fungi samples using this kit. All sizes of RNA are purified, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The procedure is rapid and convenient.

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Briefly, the cells or tissue are homogenized in a mortar and pestle using liquid nitrogen. Next, the provided Lysis Solution is added and the lysate is then spun to pellet any debris. Ethanol is then added to the clarified lysate, and the solution is loaded onto a spin-column (BIND). Under these conditions only the RNA will bind to Norgen’s resin while most of the DNA and proteins are removed in the flowthrough. The bound RNA is then washed in order to remove any remaining impurities (WASH). Lastly, the purified total RNA is eluted into the supplied Elution Solution or water (ELUTE). Please see the procedure flowchart below.

Workflow
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The RNA is preferentially purified from other cellular components, such as proteins, without the use of phenol or chloroform. The purified RNA is of the highest integrity, and 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 assays. This kit is available in two convenient sizes - 50 preps and 96 preps.


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Isolation of High Quality RNA, even from Difficult Samples. Total RNA was isolated from 50 mg samples of apple (Lanes 1), peach (Lanes 2), grape (Lanes 3), pine needle (Lanes 4), strawberry (Lanes 5) and pear (Lanes 6) using Norgen’s kit and a competitors kit. Five µL of total RNA from the 50 µL of elution was loaded on 1X MOPS 1.0 % Formaldehyde-Agarose RNA gel for analysis. Norgen’s kit allowed for the isolation of high quality RNA from all the samples, including the difficult samples, while the competitor failed to isolate RNA from grape, pine needles and strawberry. Furthermore, only Norgen’s kit was able to isolate the small RNA species (white box).


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Detection of EF1-α using one step RT-qPCR from Challenging Plant Samples. Total RNA was isolated from 50 mg samples of apple (red), peach (green), pine needle (blue) and grape leaves (burgundy) using Norgen´s Plant/Fungi Total RNA Purification Kit. Three µL of the eluted 50 µL of RNA was used in an RT-qPCR reaction for the detection of EF1-α. EF1-α was detected from all samples, indicating that the RNA is of high quality and that the Plant/Fungi RNA Purification kit is highly sensitive for total RNA isolation.


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High Yield of Plant RNA Samples isolated from Apple, Peach and Pear. Total RNA was isolated using Norgen’s kit (Blue bars) and competitor’s kit (Green bars) from 50 mg of apple, peach and pear according to the provided manual. In each case the RNA was eluted into 50 µL. RNA yield was measured by spectrophotometer in triplicate, and the average concentration was used for data. Norgen’s kit resulted in higher yields of total RNA, including miRNA. Total processing time was less with Norgen’s kit than competitor’s kit.


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High Purity of RNA Samples Isolated from Apple, Peach and Pear. RNA was isolated from 50 mg samples of apple, peach and pear using Norgen´s Plant/Fungi RNA Purification Kit and a competitor´s kit. RNA purity was determined spectrophotometrically for the RNA isolated using Norgen´s kit (260/230 = gold bars) and the competitor´s kit (260/230 = grey bars). Also shown is the 260/280 ratio for the samples. Norgen´s kit consistently isolated pure samples of Plant RNA with 260/230 ratios above 2.0, whereas the competitor´s kit had 260/230 ratios below 1.2 for the same samples.

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