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Plant Total RNA and Genomic DNA Purification Kit Add to Cart

Cat#: 24400-NB
Quantity: 50 preps
Price: 303 €
Supplier: Norgen
Shipping: RT
User Manual  

For simultaneous isolation of total RNA and DNA from the same plant sample

• High quality DNA and RNA are purified simultaneously using the same spin column
• Reduce variability - RNA and DNA are isolated from a single plant sample with no splitting of the lysate, thus reducing inconsistent results and variability.
• Isolate from small samples - Simultaneous isolation of RNA and DNA from a single sample. Ideal for precious, difficult to obtain or small samples
• All sizes of RNA are isolated, from large mRNA down to microRNA, without the use of phenol or chloroform
• Isolate DNA-free plant RNA or RNA-free plant DNA - Optional protocols for on-column DNase or RNase digestion are provided if the user wishes to isolate pure, DNA-free RNA or pure, RNA-free DNA.

Norgen’s Plant RNA/DNA Purification Kit provides a rapid method for the isolation and purification of total RNA and genomic DNA simultaneously from a single sample of plants. The total RNA and genomic DNA are both column purified in under 30 minutes using a single column. It is often necessary to isolate total RNA and genomic DNA from a single plant sample, such as for studies of gene expression, mutant or transgenic plant characterization, and host plant-pathogen characterization. Traditionally the RNA and DNA would be isolated from different aliquots of a sample, however this novel technology will allow for their simultaneous isolation from the same sample. This will not only save time, but will also be of a great benefit when isolating RNA and DNA from precious, difficult to obtain or very small samples. Furthermore, gene expression analysis will be more reliable since the RNA and DNA are derived from the same sample, therefore eliminating inconsistent results.

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Briefly, the cells or tissue of interest are lysed with the provided Lysis Solution. Next, Binding Solution is added and the lysate is spun through a provided Filter Column. Ethanol is then added to the clean lysate, and the solution is loaded onto a spin-column (BIND). Under these conditions only the RNA and DNA will bind to the column while the proteins are removed in the flowthrough. Next, an optional step can be carried out in which the genomic DNA can be digested allowing for a more pure RNA sample to be isolated. Alternatively, traces of the RNA may be digested at this point if a pure sample of genomic DNA is required instead. The bound nucleic acids are then washed in order to remove any remaining impurities (WASH). Lastly, the purified nucleic acids are eluted into the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart below.


Workflow
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Isolation of Total RNA and Genomic DNA from Tobacco, Tomato and Peach Leaf Tissue. Total RNA and genomic DNA were isolated from 50 mg of tobacco leaf, 50 mg of tomato leaf and 50 mg of peach leaf using Norgen’s Plant RNA/DNA Purification Kit. Panel A is a 1X MOPS 1.5% agarose gel showing the total RNA that was isolated after the optional on-column DNase digestion. Five µL of total RNA from each 150 µL elution was mixed with 2x RNA loading dye and denatured at 70ºC for 10 minutes and loaded onto the gel. Lane M is Norgen’s 1 kb RNA Ladder, Lanes 1 and 2 contain RNA isolated from tobacco cells, Lanes 3 and 4 contain RNA isolated from tomato cells, and Lanes 5 and 6 contain RNA isolated from peach cells. Panel B is a 1.5% agarose gel containing the genomic DNA that was isolated after the optional on-column RNase digestion, and in each case 10 µL of the 150 µL elution was loaded. Lane M is Norgen’s HighRanger 1kb DNA Ladder, Lanes 1 and 2 contain the tobacco DNA, Lanes 3 and 4 contain the tomato DNA, and Lanes 5 and 6 contain the peach DNA. The RNA and DNA are intact and of the highest quality, and can be used in a number of different downstream applications.


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Detection of 18s rDNA by Real-time PCR (SYBR Green). Total DNA and RNA from 50 mg of tobacco, tomato and peach were simultaneously isolated with Norgen’s Plant RNA/DNA Purification Kit (with the optional on-column RNase treatment). Next, 2µL of DNA from each 150 µL elution was mixed in 20 µL of PCR reaction and used in a real-time PCR (95ºC for 3min and 40 cycles at 95 ºC for 15 sec. and 60 ºC for 30 sec.). DNA from tobacco (green), tomato (red) and peach (blue) was successfully amplified, indicating the high quality of the DNA for downstream application. Primer dimer (black) also was shown.


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Isolate High Quality Total RNA from Challenging Samples. Total RNA was purified from 50 mg of grape and peach leaves using Norgen’s Plant RNA/DNA Purification Kit, with the optional on-column DNase treatment. Next, 1µL of 150 µL eluted RNA was resolved on an Agilent 2100 bioanalyzer using an RNA Nano 6000 chip. High quality total RNA was isolated from both samples, even from the challenging grape leaf sample.



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