Introduction

E.Z.N.A.^® Total RNA Kit II provides a rapid and easy method for the isolation of up to 100 ug of total RNA from tissues containing a high fat content. The kit allows single or multiple, simultaneous processing of samples in less than 30 minutes Normally 100 mg of tissue can be used in a single experiment.

While this kit may be used for isolation of RNA from whole blood, we recommend use of the E.Z.N.A.^® Blood RNA Kit (Catalog #R6614) as it is specifically designed for effective hemolysis, hemoglobin removal and provides higher RNA yields on blood, compared to this kit.
Principle

The E.Z.N.A.^® Total RNA Kit II uses the reversible binding properties of the HiBind^® matrix, a new silica-based material, with RNA-Solv^® Reagent, an Omega Bio- Inc. phenol based extraction to extract total cellular RNA from fatty tissues such as brain and adipose tissue. A specifically formulated high salt buffer system allows for more than 100 ug of RNA to bind to the matrix.

Procedure

Cells or tissues are first homogenized with RNA-Solv^® Reagent that effectively extracts the RNA from the high fat content tissue while inactivating RNases. After adding chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The aqueous phase, containing the RNA is combined with ethanol and then applied to the HiBind^® spin columns. Two separate wash steps wash away cellular debris and other contaminants. High quality RNA is then eluted in DEPC-treated sterile water.

Applications

RNA purified using the E.Z.N.A.^® Total RNA Kit II can be used in applications such as :

• RT-PCR
• Northern Blotting
• Nuclease Protection
• In vitro translation
• Poly A⁺ RNA (mRNA) purification

Formaldehyde agarose gel of total RNA purified with
E.Z.N.A.^® Total RNA Kit II. 10 ìg of total RNA isolated
from each source was loaded per line. All tissues
were from mouse.