Combined use of multiplex ligation-dependent probe amplification and automatic sequencing for identification of KAL1 defects in patients with Kallmann syndrome  Luciana Ribeiro Montenegro, Ph.D., Leticia F.G. Silveira, Ph.D., Cintia Tusset, Ph.D., Margaret de Castro, M.D., Ph.D., Beatriz R. Versiani, M.D., Ph.D., Ana Claudia Latronico, M.D., Ph.D., Berenice Bilharinho Mendonca, M.D., Ph.D., Ericka B. Trarbach, Ph.D.  Fertility and Sterility  Volume 100, Issue 3, Pages 854-859 (September 2013) DOI: 10.1016/j.fertnstert.2013.04.045 Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) MLPA results showing deletions of exons 1–2 in the case 5 with Kallmann syndrome. Normalized relative peak areas of KAL1 gene-specific and control probes are shown. Sequences present in the chromosome X in males are in single copy and have a relative peak area value of approximately 1.0. A reduction in the peak area values next to 0 indicates a deletion (black arrows). (B) PCR amplification of exons 1 (329 bp) and 2 (230 bp), showing a suspicion of a single exon 1 deletion in this same patient. Lanes 1,3 and 2,4 correspond to control male and case 5, respectively. (C) Automatic sequencing revealed that the nucleotide sequence of exon 2 amplified from patient (bottom panel) corresponding to the KALP and in the top panel, KAL1 exon 2 sequence. Fertility and Sterility 2013 100, 854-859DOI: (10.1016/j.fertnstert.2013.04.045) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Prevalence of KAL1 abnormalities in patients with Kallmann syndrome according to (A) gender and (B) type of mutation. Fertility and Sterility 2013 100, 854-859DOI: (10.1016/j.fertnstert.2013.04.045) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions