The advances in high throughput techniques for analyzing DNA and RNA have the potential to make a major impact on prevention, diagnosis, prognosis and treatment of many human cancers (1). One of the major challenges is to obtain high-quality DNA and RNA molecules. Although several RNA and DNA isolation methods for frozen tissues have been used (2-6), few efforts have been conducted to optimize these existing methods in order to isolate simultaneously high-quality RNA and DNA in an easy-to-use manner. In an effort to establish an efficient, cost-effective and simultaneous isolation of RNA and DNA from single human tumor sample, it was used RNeasy® spin columns with an unique modification of Qiagen protocol (7-8). That allows isolating simultaneously genomic DNA and totaling RNA from a single sample, with low cost and time-consuming less than 30 minutes. In contrast to other procedures where either the biological sample or the purified total nucleic acids is divided into two before being processed separately. The resulting high-quality DNA has an average length of 15-30 kb depending on homogenization and storage conditions. And it is ready to use in any downstream application, including: PCR; Southern Blot; Comparative Genome Hybridization, Methylation-specific PCR and SNP analysis.