RT Journal Article
T1 Comparison of extraction methods for the recovery, amplification and species-specific analysis of DNA from bone and bone meals
A1 Prado Rodríguez, Marta
A1 Franco Abuín, Carlos Manuel
A1 Fente Sampayo, Cristina Asunción
A1 Cepeda Sáez, Alberto
A1 Vázquez Belda, Beatriz Isabel
A1 Barros Velázquez, Jorge
K1 Bone meals
K1 DNA recovery
K1 Extraction methods
K1 Polymerase chain reaction
AB We report the effect of several parameters on the efficiency of recovery of DNA from animal bones. The effects of preheating the samples (at either 60°C or 100°C) at different intervals (from 1 h to overnight) in different media (water, 0.5 M ethylenediaminetetraacetic acid (EDTA), or 0.5 M EDTA + 0.05% sodium dodecyl sulfate (SDS) were investigated. The effect of slight (5 min) or intense (30 min) pretreatments with ultrasound was also evaluated. Several different treatments with proteinase K (ranging from 200 to 800 νg, and lasting from 1 to 3 h) at 65°C were also considered. Additionally, two different DNA extraction methods (based on silica resins and purification columns, respectively) were evaluated. The recovery of DNA from the samples was 40% higher when the bones were preheated in 0.5 M EDTA at 60°C for 1 h, this being followed by treatment with 800 νg of proteinase K for 3 h. The DNA thus obtained was successfully amplified by polymerase chain reaction (PCR) using a set of primers specific to a 359 bp region of the mitochondrial cytochrome b gene, and the species of origin were identified by visualizing the restriction fragment length polymorphism (RFLP) with the endonucleases PalI and MboI
PB Wiley
YR 2002
FD 2002-04-23
LK https://hdl.handle.net/10347/45602
UL https://hdl.handle.net/10347/45602
LA eng
NO Prado, M., Franco, C.M., Fente, C.A., Cepeda, A., Vázquez, B.I. and Barros-Velázquez, J. (2002), Comparison of extraction methods for the recovery, amplification and species-specific analysis of DNA from bone and bone meals. ELECTROPHORESIS, 23: 1005-1012. https://doi.org/10.1002/1522-2683(200204)23:7/8<1005::AID-ELPS1005>3.0.CO;2-1
NO This is the peer reviewed version of the following article: Prado, M., Franco, C.M., Fente, C.A., Cepeda, A., Vázquez, B.I. and Barros-Velázquez, J. (2002), Comparison of extraction methods for the recovery, amplification and species-specific analysis of DNA from bone and bone meals. ELECTROPHORESIS, 23: 1005-1012, which has been published in final form at https://doi.org/10.1002/1522-2683(200204)23:7/8<1005::AID-ELPS1005>3.0.CO;2-1. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited
DS Minerva
RD 24 abr 2026