Plant RNA/DNA Purification Kit

Supplier
Norgen Biotek
Cat. no.
1024400
Kit size
50
Links
Norgen Biotek Protocol Information Sheet

Features

• Complete column purification - The RNA and DNA are both column purified simultaneously using the same 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.
• 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.
• 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 DNA simultaneously from a single sample of plants. The total RNA and DNA (including genomic DNA) and 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 the same 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. The process involves first lysing the cells or tissue of interest with the provided Lysis Solution (please see the flow chart on page 4). The Lysis Solution contains detergents, as well as large amounts of a chaotropic denaturant that will rapidly inactivate any RNases that are present. A heat treatment is performed to ensure complete lysis. Ethanol is then added to the lysate, and the solution is loaded onto a spin-column. Norgen’s resin binds nucleic acids in a manner that depends on ionic concentrations, thus 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. The bound nucleic acid is then washed three times with the provided Nucleic Acid Wash Solution in order to remove any impurities, and the purified RNA and/or DNA is eluted with the Nucleic Acid Elution Buffer. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). 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. The genomic DNA is of the highest quality, and can be used in PCR reactions, sequencing, Southern blotting and SNP analysis.

Column Binding Capacity	50 µg for RNA
	15 µg for genomic DNA
Maximum Column Loading Volume	600 µL
Maximum Amount of Starting Material:
Plant Tissues	100 mg
Plant Cells	1 × 106 cells
Average Yields:
50 mg Peach Leaves	40 µg RNA, 5 µg gDNA
Size of RNA Purified	All sizes, including small RNA (<200 nt)
Time to Complete 10 Purifications	30 minutes

Isolation of Total RNA and Genomic DNA from Tobacco, Tomato and Peach Leaf Tissue

Total RNA and genomic DNA were isolated from 60 mg of tobacco leaf, 52 mg of tomato leaf and 56 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. 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. 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.

for RNA :
• qPCR
• qRT-PCR
• primer extension
• expression array assays

for DNA :
• PCR reactions
• Southern Blotting
• Sequencing
• SNP Analysis