Copyright © 2007 American Medical Association. All rights reserved. From: Direct Identification of Dermatophyte DNA From Clinical Specimens by a Nested Polymerase Chain Reaction Assay Arch Dermatol. 2007;143(6):799-816. doi:10.1001/archderm.143.6.799 Figure Legend: Representation of the ribosomal DNA (rDNA) gene complex in fungi denoting gene order and the position of the internal transcribed spacer (ITS). Primers were designed on this complex and were designated as ITS1, ITS2, ITS3, and ITS4 as their locations on genes. Oligonucleotide primers ITS1 (5′-TCC GTA GGT GAA CCT GCG G-3′) and ITS3 (5′-GGA AGT AAA AGT CGT AAC AAG G-3′) were both designed on 18S rDNA. The primer ITS2 (5′-GCT GCG TTC TTC ATC GAT GC-3′) was designed on 5.8S rDNA, and ITS4 (5′-TCC TCC GCT TAT TGA TAT GC-3′) was designed on 28S rDNA. Oligonucleotide primer pairs ITS3 and ITS4 were used in the first-round polymerase chain reaction (PCR) amplification reaction. Then the templates of nested PCR were the products of first-round PCR using ITS1 and ITS2 as paired primers. Date of download: 10/3/2017 Copyright © 2007 American Medical Association. All rights reserved.

Copyright © 2007 American Medical Association. All rights reserved. From: Direct Identification of Dermatophyte DNA From Clinical Specimens by a Nested Polymerase Chain Reaction Assay Arch Dermatol. 2007;143(6):799-816. doi:10.1001/archderm.143.6.799 Figure Legend: Genomic DNA samples extracted from 5 different skin scrapings were amplified by a MagNA pure Light Cycler System (Roche Diagnostics GmbH, Mannheim, Germany) with nested polymerase chain reaction (PCR) and analyzed by 2% agarose gel electrophoresis. The left panel represents the first round of PCR, while the right panel represents the second round. L and L′ represent the 100–base pair (bp) DNA ladder; 1 and 1′ through 5 and 5′, 5 different skin scraping tests; 6 and 6′, negative controls (Staphylococcus epidermidis genome); and 7 and 7′, positive controls (Trichophyton mentagrophytes genome). All of the distinguishable molecular weights of first PCR products were about 750 bp (lanes 5 and 7); nested PCR products, 350 bp (lanes 1′-5′ and 7′). These 5 clinical specimens belong to the genera Trichophyton and Microsporum and were proved by DNA sequencing. Bands of molecular weight that were less than 200 bp were noted in the negative control and clinical specimens (lane 1′ and lane 6′). Date of download: 10/3/2017 Copyright © 2007 American Medical Association. All rights reserved.