title: High-quality DNA from fingernails for genetic analysis.
abstract: The availability of high-quality germline DNA is an important prerequisite for a variety of genetic analyses. We have shown previously that fingernail clippings provide an optimal source of autologous, constitutional DNA for PCR-based applications. However, most existing protocols for nucleic acid purification from nails do not provide sufficiently high yields of pure and intact DNA for more demanding downstream analyses such as next generation sequencing (NGS). We have extensively tested and systematically modified a number of different protocols for DNA purification from nail material to optimize the yield and quality. The integrity of DNA was determined by PCR amplification of short (<300 bp), mid-range (>400 bp), and long-range (>2 kb) sequences using different target genes. Among the methods tested, the Prepfiler Forensic DNA Extraction kit was identified as the most appropriate approach to isolation of high-quality DNA from nail clippings. A standardized input of 20 mg nail material (1 to 10 pieces of fingernail clippings) yielded a mean of 1 μg DNA (range, 0.5 to 2.3 μg). Subsequent PCR-analysis revealed efficient amplifiability of short and mid-range targets in 93% and 90%, and long-range fragments in 60% of the samples tested. The adequacy for next generation sequencing applications was demonstrated by successful high-resolution HLA-typing in ten transplant recipients. Hence, the protocol presented facilitates the exploitation of fingernail material even for demanding genomic analyses both in research and diagnostics.
authors.affiliation: Children's Cancer Research Institute, Vienna, Austria; Labdia Labordiagnostik GmbH, Vienna, Austria.
authors.forename: Sandra
authors.initials: S
authors.lastname: Preuner
authors.affiliation: Red Cross Transfusion Service for Upper Austria, Linz, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Danzer
authors.affiliation: Red Cross Transfusion Service for Upper Austria, Linz, Austria.
authors.forename: Johannes
authors.initials: J
authors.lastname: Pröll
authors.affiliation: Children's Cancer Research Institute, Vienna, Austria.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Pötschger
authors.affiliation: St. Anna Children's Hospital, Vienna, Austria.
authors.forename: Anita
authors.initials: A
authors.lastname: Lawitschka
authors.affiliation: Red Cross Transfusion Service for Upper Austria, Linz, Austria.
authors.forename: Christian
authors.initials: C
authors.lastname: Gabriel
authors.affiliation: Children's Cancer Research Institute, Vienna, Austria; Labdia Labordiagnostik GmbH, Vienna, Austria; Department of Pediatrics, Medical University of Vienna, Vienna, Austria. Electronic address: thomas.lion@ccri.at.
authors.forename: Thomas
authors.initials: T
authors.lastname: Lion
mesh_terms: D004247:DNA / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D005820:Genetic Testing; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006650:Histocompatibility Testing / Q000379:methods; D006801:Humans; D009262:Nails / Q000737:chemistry / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004247:DNA