Abstract

AMARU, Ricardo et al. UMSAgen, método para la extracción simultánea de RNA y DNA para diagnóstico molecular. Cuad. - Hosp. Clín. [online]. 2008, vol.53, n.1, pp.38-43. ISSN 1562-6776.

INTRODUCTION The RNA and the DNA are genetic materials of the cell. The RNA/DNA extraction is necessary in modern medicine. Several RNA/DNA isolation methods exist but they are very expensive and therefore not applicable in our country. In this work, we describe the UMSAgen method as an alternative for simultaneous total RNA and genomic DNA extraction for molecular diagnosis. MATERIAL AND METHODS Twenty bone marrow samples of chronic myeloid leukemia patients were obtained. The leukocytes were separated from the erythrocytes with lyses solution; 3x106 cells were lysed in guanidio solution and conserved at -20ºC until the extraction procedure. The extraction of nucleotide acids was realized by 3 different methods: a) commercial method QuiagenTM for RNA extraction, b) classic method (phenol/chloroform) for DNA extraction and c) UMSAgen method for simultaneous RNA/DNA extraction. The two first methods were used as a control test for UMSAgen. RESULTS The quality and quantity of total RNA and genomic DNA obtained by the UMSAgen method were similar to that of the QuiagenTM Kit and the classic method, respectively. The ABL-BCR and JAK-2V617F amplification by RT-PCR and PCR were successful. CONCLUSIONS The UMSAgen is an easy and inexpensive altrernative method for total RNA and genomic DNA extraction in diagnostic and biomolecular research.

Keywords : RNA; DNA; extraction; PCR; RT- PCR and electrophoresis.

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