RNA Extraction/Total RNA
The Presto™ miRNA Purification Kit is designed for purifying and enriching of small RNA molecules (< 200 nt) from a wide variety of samples, such as animal tissues, cultured cells and bacterial cells. These small RNAs include microRNA (miRNA), short interfering RNA (siRNA), as well as tRNA and 5S ribosomal RNA. miRNAs and siRNAs are typically 20-25 nucleotides long and vital for regulating gene expression by binding to mRNA molecules and affecting their stability or translation. Purification is based on spin column technology without the use of phenol and chloroform. Briefly, the cells or tissues are lysed using detergents and chaotropic salt. The larger RNA fragments in the chaotropic salt are bound by the glass fiber matrix of the first spin column while the smaller RNA molecules pass through into the flow-through. The smaller RNA molecules in the flow-through are bound by the second spin column once any contaminants have been removed using the Wash Buffer. The small RNA molecules are then eluted using the RNase-free water, and are ready to be used in various applications, including RT-PCR, northern blotting and microarray analysis.