Ciara Rhodes PhD, Reilly Price MS, Celeste Willetts BS, Jenise Swall PhD, Laura Gaydosh Combs PhD, Sarah Seashols-Williams PhD
{"title":"利用脱矿化骨片改善风化牛骨的DNA恢复和STR谱发展。","authors":"Ciara Rhodes PhD, Reilly Price MS, Celeste Willetts BS, Jenise Swall PhD, Laura Gaydosh Combs PhD, Sarah Seashols-Williams PhD","doi":"10.1111/1556-4029.70023","DOIUrl":null,"url":null,"abstract":"<p>Current processing techniques for harvesting DNA from osseous elements are destructive, and success rates vary widely. When skeletal elements are homogenized into a fine powder, endogenous DNA may be subjected to fragmentation, and the likelihood of introducing co-purified inhibitory substances to the sample increases. While a limited number of articles in the relevant literature have challenged the status quo of pulverization, powdering hard tissue samples before DNA isolation continues to be standard practice in the forensic and ancient DNA communities. In this work, we have developed and optimized an alternative front-end processing method for demineralizing and slicing cortical bone using aged and weathered bovine skeletal samples as a model for degraded human bone. Additionally, this study evaluated the enzymatic digestion of demineralized bone slices using collagenase I as a powder-free alternative for cell isolation. The efficacy of pulverization, demineralized slices, and collagenase-digested demineralized slices was assessed via DNA quantitation and STR profile data. The combined treatment incorporating collagenase digestion of demineralized slices did not improve the recovery of endogenous DNA yields or STR profile development; however, profiles developed from demineralized slices retained a significantly greater percentage of expected alleles and higher peak heights than samples processed with conventional powdering and organic chemistry extraction. By eliminating pulverization of bone and employing modified silica-based extraction chemistry, the susceptibility to inhibitory and competitive effects from native and foreign components often found in degraded skeletal remains can be minimized when utilizing total demineralization of small fragments with subsequent slicing before lysis.</p>","PeriodicalId":15743,"journal":{"name":"Journal of forensic sciences","volume":"70 3","pages":"954-963"},"PeriodicalIF":1.5000,"publicationDate":"2025-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1556-4029.70023","citationCount":"0","resultStr":"{\"title\":\"Improved DNA recovery and STR profile development from weathered Bos taurus bones using demineralized bone slices\",\"authors\":\"Ciara Rhodes PhD, Reilly Price MS, Celeste Willetts BS, Jenise Swall PhD, Laura Gaydosh Combs PhD, Sarah Seashols-Williams PhD\",\"doi\":\"10.1111/1556-4029.70023\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Current processing techniques for harvesting DNA from osseous elements are destructive, and success rates vary widely. When skeletal elements are homogenized into a fine powder, endogenous DNA may be subjected to fragmentation, and the likelihood of introducing co-purified inhibitory substances to the sample increases. While a limited number of articles in the relevant literature have challenged the status quo of pulverization, powdering hard tissue samples before DNA isolation continues to be standard practice in the forensic and ancient DNA communities. In this work, we have developed and optimized an alternative front-end processing method for demineralizing and slicing cortical bone using aged and weathered bovine skeletal samples as a model for degraded human bone. Additionally, this study evaluated the enzymatic digestion of demineralized bone slices using collagenase I as a powder-free alternative for cell isolation. The efficacy of pulverization, demineralized slices, and collagenase-digested demineralized slices was assessed via DNA quantitation and STR profile data. The combined treatment incorporating collagenase digestion of demineralized slices did not improve the recovery of endogenous DNA yields or STR profile development; however, profiles developed from demineralized slices retained a significantly greater percentage of expected alleles and higher peak heights than samples processed with conventional powdering and organic chemistry extraction. By eliminating pulverization of bone and employing modified silica-based extraction chemistry, the susceptibility to inhibitory and competitive effects from native and foreign components often found in degraded skeletal remains can be minimized when utilizing total demineralization of small fragments with subsequent slicing before lysis.</p>\",\"PeriodicalId\":15743,\"journal\":{\"name\":\"Journal of forensic sciences\",\"volume\":\"70 3\",\"pages\":\"954-963\"},\"PeriodicalIF\":1.5000,\"publicationDate\":\"2025-03-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1556-4029.70023\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of forensic sciences\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/1556-4029.70023\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MEDICINE, LEGAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of forensic sciences","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/1556-4029.70023","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MEDICINE, LEGAL","Score":null,"Total":0}
Improved DNA recovery and STR profile development from weathered Bos taurus bones using demineralized bone slices
Current processing techniques for harvesting DNA from osseous elements are destructive, and success rates vary widely. When skeletal elements are homogenized into a fine powder, endogenous DNA may be subjected to fragmentation, and the likelihood of introducing co-purified inhibitory substances to the sample increases. While a limited number of articles in the relevant literature have challenged the status quo of pulverization, powdering hard tissue samples before DNA isolation continues to be standard practice in the forensic and ancient DNA communities. In this work, we have developed and optimized an alternative front-end processing method for demineralizing and slicing cortical bone using aged and weathered bovine skeletal samples as a model for degraded human bone. Additionally, this study evaluated the enzymatic digestion of demineralized bone slices using collagenase I as a powder-free alternative for cell isolation. The efficacy of pulverization, demineralized slices, and collagenase-digested demineralized slices was assessed via DNA quantitation and STR profile data. The combined treatment incorporating collagenase digestion of demineralized slices did not improve the recovery of endogenous DNA yields or STR profile development; however, profiles developed from demineralized slices retained a significantly greater percentage of expected alleles and higher peak heights than samples processed with conventional powdering and organic chemistry extraction. By eliminating pulverization of bone and employing modified silica-based extraction chemistry, the susceptibility to inhibitory and competitive effects from native and foreign components often found in degraded skeletal remains can be minimized when utilizing total demineralization of small fragments with subsequent slicing before lysis.
期刊介绍:
The Journal of Forensic Sciences (JFS) is the official publication of the American Academy of Forensic Sciences (AAFS). It is devoted to the publication of original investigations, observations, scholarly inquiries and reviews in various branches of the forensic sciences. These include anthropology, criminalistics, digital and multimedia sciences, engineering and applied sciences, pathology/biology, psychiatry and behavioral science, jurisprudence, odontology, questioned documents, and toxicology. Similar submissions dealing with forensic aspects of other sciences and the social sciences are also accepted, as are submissions dealing with scientifically sound emerging science disciplines. The content and/or views expressed in the JFS are not necessarily those of the AAFS, the JFS Editorial Board, the organizations with which authors are affiliated, or the publisher of JFS. All manuscript submissions are double-blind peer-reviewed.