01962naa a2200145 a 450000100080000000500110000800800410001910000230006024502060008326000090028952013120029870000160161070000200162677301700164614711732009-02-17       2008    bl ---       0--  u    #d1 aSOSA-GÓMEZ, D. R.  aNuevo método de PCR para estudiar Ia región espaciadora intergénica de Nomuraea rileyi (Farlow) Samson. New PCR method to study the intergenic spacer (IGS) region of Nomuraea rileyi (Farlow) Samson.  c2008  aThe variability in the IGS region of the rDNA gene complex has proven useful in the development of PCR primers for strain identification. However, the lack of information on this region in Nomuraea rileyi has precluded the development of specific primers for PCR amplification. Also, due to the high variability in this region, amplification is not possibIe with universal primers. Thus, to obtain sequences of the of the  region a new approach was used, a combination of IGS primer with 10-mer oligonucleotides (OD11 or EO4, Operon Technologies) were used. The amplification products were visualized by agarose gel electrophoresis. DNA bands were selected for comparison between IGS-10 mer primer product amplification and DNA bands produced by 10-mer amplification. When DNA products were absent in the RAPD amplification and present in the combination IGS-10 mer amplification, these products were gelpurified, ligated into the plasmid vector TOPO TA (lnvitrogen), and amplified in DH10 B Escherichia coli cells. The DNA obtained from positive clones were subjected to Sanger sequencing. The obtained sequences ranged from 1,027 to 1,064 bases. Comparisons among isolates obtained from the same geographic region suggest that this technique could be used to develop strain specific primers for N. rileyi.1 aBINNECK, E.1 aMARIN, S. R. R.  tIn: CONGRESO LATINOAMERICANO DE MICOLOGIA, 6., 2008, Mar del Plata. Libro de resúmenes. Buenos Aires: Asociación Latinoamericana de Micologia, 2008. p. III. Anexo.