Bioteke’s General Plant RNA 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.
Bioteke’s General Plant RNA 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 solution chromatography using Bioteke’s proprietary resin as the separation matrix. Bioteke’s resin binds RNA in a manner that depends on ionic concentrations. The small RNA molecules are preferentially purified from other cellular components such as ribosomal RNA without the use of phenol or chloroform. The process involves the use of two different spin columns: the Large RNA Removal Columnand the microRNA Enrichment Column (please see flow chart on page 3). Briefly, the cells or tissues of interest are lysed using the provided Lysis Buffer C, the lysate is spun through the provided the Filter Column, and then ethanol is added to the clarified lysate. The lysate is then applied to the Large RNA Removal Column, and the larger RNA molecules will bind to the resin in the spin column while the smaller RNA species will pass through into the flowthrough. Ethanol is then added to the flowthrough, and the sample is applied to the microRNA Enrichment Column. The small RNA molecules will then bind to the resin, and any impurities are removed through a series of washes with the provided Wash Solution A. The small RNA molecules are then eluted using the Elution Solution, and are ready for use in various applications.
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.