During the second round of PCR primers, forward inner HB F2 primer 1228-1247 5'-ATCAGCGCATGCGTGGAACC-3 and reverse inner HB R2 1854-1877 5'-TGGAGGCTG AACAGTGGGACAT-3' were used with the same cycling conditions as in the first round except that alignment was at 61°C. This was amplifed to 649 base pairs.
Durante la segunda ronda de PCR se usaron los primers internos forward HB-Inner F2 1228-1247 5'- ATCAGCGCATGCGTGGAACC-3' y reverse HB-Inner R2 1854-1877 5'- TGGAGGCTGA ACAGTGGGACAT-3' y se siguieron las mismas condiciones de ciclado de la primera ronda, a excepción del alineamiento a 61 °C, cuyo amplificado fue de 649 pares de bases.