Extraction of DNA from trace forensic samples with a modified lysis buffer and chitosan coated magnetic beads

doi:10.1016/j.fsigen.2023.102932

	Extraction of DNA from trace forensic samples with a modified lysis buffer and chitosan coated magnetic beads
	Hu, Anzhong 2,3; Chen, Lin 1; Geng, Xuelei 1; Zhu, Ling 2; Liu, Yong 2; Yang, Ke 2; Zhu, Huaqing 3; Zhu, Cancan 2
刊名	FORENSIC SCIENCE INTERNATIONAL-GENETICS
	2023-11-01
卷号	67
关键词	Chitosan@Beads Forensic biological evidence STR loci Trace DNA extraction Inhibitors Polymerase chain reaction (PCR) Short tandem repeat (STR)
ISSN号	1872-4973
DOI	10.1016/j.fsigen.2023.102932
通讯作者	Zhu, Huaqing(zhuhq@ahmu.edu.cn) ; Zhu, Cancan(zhucancan@aiofm.ac.cn)
英文摘要	The trace amounts of human tissue cells or body fluids left at the crime scene are often mixed with inhibitors such as rust, pigments, and humic acid. The extraction of the DNA from the trace cells is crucial for the investigation of cases. Usually, specially designed magnetic nanoparticles were chosen by the case investigators to enrich and elute DNA, which was then used for polymerase chain reaction (PCR) and short tandem repeat (STR) analysis. The traditional approach often had the following problems, such as low DNA enrichment efficiency, possible DNA breakage, and complex operations. Here, the 1%(w/v) of chitosan (75% deacetylation degree) was used to modify the 50 nm magnetic nanoparticles to gain the Chitosan@Beads, which theoretically carried positively charged in the pH = 5 of lysis buffer so as to adsorb negatively charged DNA through electrostatic interactions. The XPS and FT-IR results demonstrated that chitosan was successfully attached to the surface of magnetic nanoparticles. A set of simulated samples, containing 20 mg/mu L of humic acid, pigments, iron ions (Fe2+, Fe3+), and the coexistence of the above three substances, were prepared to simulate the case scene. Human bronchial epithelial cells were mixed with the 200 mu L of the above simulated samples for DNA extraction. 400 mu L of lysis buffer, 20 mu L of proteinase K (10 mg/mL) and 20 mu L of Chitosan@Beads solution (20 mg/mL) was used for cell disruption and DNA enrichment. The extraction sensitivity of Chitosan@Beads was confirmed to be 10 cells, superior to commercial reagent kits. The Chitosan@Beads@DNA can directly use for "In-situ PCR" with elutionfree operations. The STR loci rate of DNA extracted by Chitosan@Beads was around 97.9%, higher than the commercial kit (66.7%). In short, we foresee here developed novel Chitosan@Beads and modified lysis buffer could provide a new model for the DNA extraction of forensic trace evidence.
资助项目	National Natural Science Founda- tion of China[82002189] ; Key Research and Development Program of Anhui Province of China[202004d07020014] ; National Key Research and Development Program of China[2022YFC2302700]
WOS研究方向	Genetics & Heredity ; Legal Medicine
语种	英语
出版者	ELSEVIER IRELAND LTD
WOS记录号	WOS:001078556200001
资助机构	National Natural Science Founda- tion of China ; Key Research and Development Program of Anhui Province of China ; National Key Research and Development Program of China
内容类型	期刊论文
源URL	[http://ir.hfcas.ac.cn:8080/handle/334002/132513]
专题	中国科学院合肥物质科学研究院
通讯作者	Zhu, Huaqing; Zhu, Cancan
作者单位	1.Dept Anhui Publ Secur, Inst Forens Sci, Hefei 230061, Peoples R China 2.Chinese Acad Sci, Anhui Inst Opt & Fine Mech, Hefei Inst Phys Sci, Hefei 230031, Peoples R China 3.Anhui Med Univ, Sch Basic Med, Hefei 230032, Peoples R China
推荐引用方式 GB/T 7714	Hu, Anzhong,Chen, Lin,Geng, Xuelei,et al. Extraction of DNA from trace forensic samples with a modified lysis buffer and chitosan coated magnetic beads[J]. FORENSIC SCIENCE INTERNATIONAL-GENETICS,2023,67.
APA	Hu, Anzhong.,Chen, Lin.,Geng, Xuelei.,Zhu, Ling.,Liu, Yong.,...&Zhu, Cancan.(2023).Extraction of DNA from trace forensic samples with a modified lysis buffer and chitosan coated magnetic beads.FORENSIC SCIENCE INTERNATIONAL-GENETICS,67.
MLA	Hu, Anzhong,et al."Extraction of DNA from trace forensic samples with a modified lysis buffer and chitosan coated magnetic beads".FORENSIC SCIENCE INTERNATIONAL-GENETICS 67(2023).