RNA PURIFICATION OVERVIEW
The quality of RNA obtained from biological samples or reaction mixtures is of critical importance to the success of downstream applications including real-time quantitative PCR, reverse transcription PCR, microarrays, Northern blot analysis, nuclease protection assays, RNA mapping, in vitro translation, cDNA library construction, etc.
Thus, it is essential that the methods of extracting, purifying or isolating RNA from different types of cells and tissues produce RNA of sufficient yield and structural integrity. With this in mind, the scientists at Zymo Research have developed a range of RNA extraction, purification and isolation kits designed for the efficient recovery of high quality RNA from a diversity of sources.
Purification of RNA from different types of cells and tissues has been difficult in the past, however high quality RNA is essential for the success of downstream applications such as microarrays, RNA transfection, denaturing-gel electrophoresis, Northern blotting and RT-PCR. Zymo Research Corporation has used their state of the art ‘Fast-Spin’ column technology to produce the RNA Clean & Concentrator kit. This kit recovers a highly concentrated RNA, while still ensuring the structural integrity of the product.
Zymo Research offers an assortment of products that allow for the simple, rapid, and efficient isolation of total RNA from a variety of biological sources including fresh, frozen, or paraffin-embedded tissues, cultured cells, buccal cells, whole blood, plasma, serum, urine, yeast, or RNA viruses. Like our RNA clean-up kits, all of the RNA isolation kits feature Fast-Spin column technology for highly concentrated RNA that is well suited for applications like microarrays, denaturing-gel electrophoresis, Northern blotting, and RT-PCR. The versatile solution for total RNA purification, Quick-RNA kits are available in Micro-, Mini-, Midi- and 96-well format. Additionally, the ZR RNA MicroPrep™ and ZR RNA MiniPrep™ kits are capable of isolating RNA species as small as 17 nt. Each kit has been optimized for a particular application with specialized, nuclease-free components that ensure: 1) maximum levels of membrane solubilization and cellular disruption, 2) total inhibition of nuclease activity, 3) complete deproteinization of the sample, 4) efficient isolation and concentration of the RNA, 5) stabilization and safe storage of the RNA.
Agarose and polyacrylamide gel electrophoresis are two common techniques for determining the size, purity, and sequence identity (e.g., Northern and Southern blotting procedures) of nucleic acids. Nucleic acids can be visualized in gels using fluorescent stains (e.g. ethidium bromide) or by silver staining. RNA size markers (or ladders) are necessary to help identify unknown nucleic acids in gel-based procedures. The ZR small-RNA™ Ladder is ideal for identifying small RNAs (i.e., siRNAs and miRNAs) from 17 - 29 bases in size.