The E.Z.N.A.® Total RNA Kit II is designed for isolating total cellular RNA from tissues rich in fat, such as brain adipose tissues. However, this kit can also be used for the isolation of total RNA from other type of tissues including cultured eukaryotic cells, animal tissues, or bacteria.
The E.Z.N.A.® Total RNA Kit II uses the reversible binding properties of HiBind® matrix, a new silica-based material. By combining the high lysis efficiency of RNA-Solv® Reagent with Omega Bio-tek’s innovative HiBind® technology, this kit can extract total cellular RNA from all types of animal or human tissues including fatty tissues such as brain and adipose tissue. A specifically formulated high salt buffer system allows more than 100 µg RNA molecules greater than 200 bases to bind to the matrix. Cells or tissues are first homogenized with RNA-Solv® Reagent that inactivates RNases. After adding chloroform, the homogenate is separated into aqueous and organic phase by centrifugation. The aqueous phase which contains the RNA is adjusted with ethanol and applied to the HiBind® RNA Mini Column to which total RNA binds, while cellular debris and other contaminants are washed away. High-quality RNA is eluted in DEPC Water.
For Research Use Only. Not for use in diagnostic procedures.
FEATURES | SPECIFICATIONS |
---|---|
Downstream Application | PCR, qPCR, real-time RT-PCR, microarray, Northern blot, poly-A purification |
Elution Volume | 45-75 μL |
Starting Material | Tissues rich in fat |
Starting Amount | 1 x 107 eukaryotic cells or 25-30 mg tissue |
RNA Yield | Up to 100 µg |
Processing Mode | Manual, centrifugation/vacuum |
Throughput | 1 - 24 |
RNA Binding Technology | Silica Mini spin column |
Binding Capacity | 100 µg |
Figure 1. Total RNA purified using the E.Z.N.A.® Total RNA Kit II. Purified total RNA from mouse tissue samples was isolated with the E.Z.N.A.® Total RNA Kit II. Total RNA (300 ng/lane) was analyzed on a 1% agarose gel to demonstrate yield and quality of the RNA. M: CL5000 DNA Marker.
Figure 2. RNA was isolated from 10 mg chicken liver using the E.Z.N.A.® Total RNA Kit II. A serial dilution of recovered RNA was then used in a SYBR® Green ral-time RT-PCR targeting a 150 bp fragment of the β-actin. Each reaction was performed in 40 replicates. The lines represent input RNA amounts of 50, 5, 0.5, 0.05 and 0.005 ng (left to right) per reaction.
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