( d WBCs make up only about 0.2% of blood...)Improve RNA Isolation for Array Analysis and More

d WBCs make up only about 0.2% of blood cells and 2) whole blood also contains large amounts of proteins, heme, and genomic DNA, as well as intracellular and extracellular RNases. Thus, WBCs typically need to be isolated away from the rest of the blood constituents prior to RNA isolation.

The RiboPure-WBC Kit protocol consists of three steps: 1) fractionation of whole blood by centrifugation, recovery of the WBCs and RNA stabilization in Ambion's RNAlater (provided), 2) lysis of WBCs in guanidinium and initial RNA purification by phenol/chloroform extraction, and 3) final RNA purification on a glass fiber filter.

The kit provides sufficient reagents for the purification and stabilization of WBCs from 40 whole blood samples of up to 10 ml each (2.5-10 ml of whole blood can be used per isolation). The RNA purified with the RiboPure-WBC Kit can be used in the most demanding downstream applications, such as real-time RT-PCR or array analysis. Average yields of RNA range between 4-20 g from 5 ml of whole blood; the variation is due to differences in WBC numbers between samples.

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