AIM: To quantitatively compare a silica extraction method with a commonly used phenol/chloroform extraction method for DNA analysis of specimens exhumed from mass graves. METHODS: DNA was extracted from twenty randomly chosen femur samples, using the International Commission on Missing Persons (ICMP) silica method, based on Qiagen Blood Maxi Kit, and compared with the DNA extracted by the standard phenol/chloroform-based method. The efficacy of extraction methods was compared by real time polymerase chain reaction (PCR) to measure DNA quantity and the presence of inhibitors and by amplification with the PowerPlex 16 (PP16) multiplex nuclear short tandem repeat (STR) kit. RESULTS: DNA quantification results showed that the silica-based method extracted on average 1.94 ng of DNA per gram of bone (range 0.25-9.58 ng/g), compared with only 0.68 ng/g by the organic method extracted (range 0.0016-4.4880 ng/g). Inhibition tests showed that there were on average significantly lower levels of PCR inhibitors in DNA isolated by the organic method. When amplified with PP16, all samples extracted by silica-based method produced 16 full loci profiles, while only 75% of the DNA extracts obtained by organic technique amplified 16 loci profiles. CONCLUSIONS: The silica-based extraction method showed better results in nuclear STR typing from degraded bone samples than a commonly used phenol/chloroform method.
AIM: To quantitatively compare a silica extraction method with a commonly used phenol/chloroform extraction method for DNA analysis of specimens exhumed from mass graves. METHODS: DNA was extracted from twenty randomly chosen femur samples, using the International Commission on Missing Persons (ICMP) silica method, based on Qiagen Blood Maxi Kit, and compared with the DNA extracted by the standard phenol/chloroform-based method. The efficacy of extraction methods was compared by real time polymerase chain reaction (PCR) to measure DNA quantity and the presence of inhibitors and by amplification with the PowerPlex 16 (PP16) multiplex nuclear short tandem repeat (STR) kit. RESULTS: DNA quantification results showed that the silica-based method extracted on average 1.94 ng of DNA per gram of bone (range 0.25-9.58 ng/g), compared with only 0.68 ng/g by the organic method extracted (range 0.0016-4.4880 ng/g). Inhibition tests showed that there were on average significantly lower levels of PCR inhibitors in DNA isolated by the organic method. When amplified with PP16, all samples extracted by silica-based method produced 16 full loci profiles, while only 75% of the DNA extracts obtained by organic technique amplified 16 loci profiles. CONCLUSIONS: The silica-based extraction method showed better results in nuclear STR typing from degraded bone samples than a commonly used phenol/chloroform method.
Authors: A Alonso; S Andelinović; P Martín; D Sutlović; I Erceg; E Huffine; L F de Simón; C Albarrán; M Definis-Gojanović; A Fernández-Rodriguez; P García; I Drmić; B Rezić; S Kuret; M Sancho; D Primorac Journal: Croat Med J Date: 2001-06 Impact factor: 1.351
Authors: Antonio Alonso; Pablo Martin; Cristina Albarrán; Pilar Garcia; Lourdes Fernandez de Simon; Maria Jesús Iturralde; Amparo Fernández-Rodriguez; Inmaculada Atienza; Javier Capilla; Julia García-Hirschfeld; Pilar Martinez; Gloria Vallejo; Oscar García; Emilio García; Pilar Real; David Alvarez; Antonio León; Manuel Sancho Journal: Croat Med J Date: 2005-08 Impact factor: 1.351
Authors: J P Whitaker; T M Clayton; A J Urquhart; E S Millican; T J Downes; C P Kimpton; P Gill Journal: Biotechniques Date: 1995-04 Impact factor: 1.993
Authors: Ana Milos; Arijana Selmanović; Lejla Smajlović; René L M Huel; Cheryl Katzmarzyk; Adi Rizvić; Thomas J Parsons Journal: Croat Med J Date: 2007-08 Impact factor: 1.351