Omega Bio-Tek Fungal RNA Mini Kit
(Homogenizer Columns Included)
5 preps With V-Spin column
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Isolation of cellular RNA from Fungal cells or tissue
Modified from the E.Z.N.A.® Plant RNA Kit, the E.Z.N.A.® Fungal RNA Kit uses an improved buffer system that is more efficient for fungal samples. This kit provides convenient and rapid methods for the isolation of total RNA from a variety of fungal species. Rather, the method involves a simple and rapid precipitation step for removal of much of the polysaccharides and phenolic compounds commonly found in fungal tissues. In combination with HiBind ® RNA spin columns, this method permits purification of high-quality RNA from as much as 200 mg tissue. The system is also efficient enough to purify total RNA from as little as 10 mg of tissue or 100 cells. Purified RNA can be used for many downstream applications such as RT-PCR, Northern Blotting, Poly A +(mRNA) purification, and differential display.
The Omega Biotek Fungal RNA Isolation Mini Kit provides a convenient and rapid method for the isolation of total RNA from a variety of plant samples. This kit are ideal for processing multiple samples in less than an hour. This kit does not require the use of cumbersome or expensive shredding/homogenizing accessories as an attempt to shear genomic DNA in viscous cell lysates. Rather, the method involves a simple and rapid precipitation step for removal of much of the polysaccharides and phenolic compounds commonly found in such tissues. In combination with our proprietary HighBind RNA EZ-SpinKit columns and an optimized buffer system, this procedure permits purification of high quality RNA from fresh or frozen fungi (including liquid nitrogen frozen samples). This mini-sized kit can isolate as much as 100mg of fungal tissue and as little as 10mg of tissue or 100 cells in less than 30 minutes. Yield is 30 60μg. This kit can also be used to purify RNA from enzymatic reaction mixtures and to desalt RNA. Purified RNA has O.D. 260/O.D.280 ratios of 1.8 2.0 and is suitable for downstream applications such as RT-PCR, Northern analysis, differential display, and poly A+ RNA selection. An improved protocol is included for optional on-column DNase digestion procedure. This kit eliminates the need for for phenol/chloroform extractions and CSCl gradient ultracentrifugation and precipitation with isopropanol.
Yields obtained with the Fungal Total RNA Isolation EZ-SpinKit: Acremonium - 50μg; Fusarium avenaceu - 37μg; Mushrooms - 43μg
Total RNA purified using the E.Z.N.A.® Fungal RNA Kit. Purified total RNA from 100 mg fungal species was isolated with the E.Z.N.A.® Fungal RNA Kit. RNA was extracted from fungal species as follows: sample 1 from E. edodes, sample 2 from V. volvacea, sample 3 from F. velutipes, sample 4 from H. erinaceum. Total RNA (10% of total purified RNA) was analyzed to demonstrate yield and quality of the RNA.
RNA Isolation and Purification
Manipulation of RNA is inherently more challenging than that of DNA since RNase is not only ubiquitous to most laboratories, it is also extremely unstable. Omega RNA EZ-Kits and EZ-96 plate Kits allow for reliable isolation of RNA from a variety of sources with its HighBind spin column method. These systems involve the simultaneous lysis of cells from a variety of sources of endogenous RNases. This is accomplished by using proprietary denaturing buffers that enhance the reversible binding of RNA to the HighBind silica matrix. While cells can be lysed directly, tissues are disrupted or homogenized using common laboratory supplies. Binding conditions are further adjusted and lysates are applied to the spin columns that selectively bind RNA molecules greater than 200 nt in length, thus favoring mRNA isolation. Successive washes with specially formulated buffers remove inhibitors and contaminants, allowing subsequent elution of pure RNA with water. As with other Omega products, phenol and chloroform extraction are eliminated. Kits are tailor made for specific cells and tissue sources to ensure consistent and reliable RNA preps, every time.