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Download Catalog 2009/2010

Ron´s Blood RNA Mini Kit

Manual for Ron´s Blood RNA Mini Kit

Introduction

Ron´s Blood RNA Kits are designed for isolation of total intracellular RNA from up to 1.5 ml of fresh, whole blood treated with any common anticoagulant such as heparin, EDTA, or acid-citrate-dextrose. One ml of blood typically yields 1 – 5 µg of total RNA. The procedure completely removes contaminants and enzyme inhibitors making total RNA isolation fast, less than 60 min. needed, convenient, and reliable. There is no need for phenol/chloroform extractions, and time-consuming steps such as CsCl gradient ultracentrifugation, and precipitation with isopropanol or LiCl, are eliminated.

RNA purified using the Ron´s Blood RNA method is ready for applications such as RT-PCR, Poly(A)⁺-RNA extractions, Nor­thern Blots, Nuclease Protection Assays and <em>in vitro</em> translations.

Principle

The Ron´s Blood RNA Kits use the reversible binding properties of PerfectBind matrix, a new silica-based material. This is combined with the speed of mini-column spin technology. Red blood cells are selectively lysed and white cells collected by centrifugation. After lysis of white blood cells under denaturing conditions that inactivate RNases, total RNA is purified in the PerfectBind spin column. A specially formulated high salt buffer system allows RNA molecules greater than 200 bases to bind to the matrix. Cellular debris and other contaminants (such as hemoglobin) are effectively washed away and high quality RNA is finally eluted in RNase-free water.

Ron´s Blood RNA Kits are specially designed for isolation of total RNA out of 1.5 ml maximum sample volume. The functionality of the kit is not restricted by the binding capacity of the PerfectBind RNA Column, but it is restricted by the maximum sample volume. Due to the large quantity of erythrocytes and proteins in the blood, yield and quality of the received RNA are reduced clearly on use of larger sample volumes.

It is also possible to use this kit for RNA isolation out of frozen blood sample, but the sample volume is then restricted to 150 µl for each column.

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