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Assessment of DNA quality for whole genome library preparation.
Jansson, Linda; Aili Fagerholm, Siri; Börkén, Emelie; Hedén Gynnå, Arvid; Sidstedt, Maja; Forsberg, Christina; Ansell, Ricky; Hedman, Johannes; Tillmar, Andreas.
Afiliação
  • Jansson L; National Forensic Centre, Swedish Police Authority, Linköping, Sweden; Applied Microbiology, Department of Chemistry, Lund University, Lund, Sweden.
  • Aili Fagerholm S; National Forensic Centre, Swedish Police Authority, Linköping, Sweden.
  • Börkén E; Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Linköping, Sweden.
  • Hedén Gynnå A; National Forensic Centre, Swedish Police Authority, Linköping, Sweden.
  • Sidstedt M; National Forensic Centre, Swedish Police Authority, Linköping, Sweden.
  • Forsberg C; National Forensic Centre, Swedish Police Authority, Linköping, Sweden.
  • Ansell R; National Forensic Centre, Swedish Police Authority, Linköping, Sweden; Department of Physics, Chemistry and Biology, IFM, Linköping University, Linköping, Sweden.
  • Hedman J; National Forensic Centre, Swedish Police Authority, Linköping, Sweden; Applied Microbiology, Department of Chemistry, Lund University, Lund, Sweden.
  • Tillmar A; Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Linköping, Sweden; Department of Biomedical and Clinical Sciences, Faculty of Medicine and Health Sciences, Linköping University, Linköping, Sweden. Electronic address: andreas.tillmar@liu.se.
Anal Biochem ; 695: 115636, 2024 Dec.
Article em En | MEDLINE | ID: mdl-39111682
ABSTRACT
In recent years, more sophisticated DNA technologies for genotyping have enabled considerable progress in various fields such as clinical genetics, archaeogenetics and forensic genetics. DNA samples previously rejected as too challenging to analyze due to low amounts of degraded DNA can now provide useful information. To increase the chances of success with the new methodologies, it is crucial to know the fragment size of the template DNA molecules, and whether the DNA in a sample is mostly single or double stranded. With this knowledge, an appropriate library preparation method can be chosen, and the DNA shearing parameters of the protocol can be adjusted to the DNA fragment size in the sample. In this study, we first developed and evaluated a user-friendly fluorometry-based protocol for estimation of DNA strandedness. We also evaluated different capillary electrophoresis methods for estimation of DNA fragmentation levels. Next, we applied the developed methodologies to a broad variety of DNA samples processed with different DNA extraction protocols. Our findings show that both the applied DNA extraction method and the sample type affect the DNA strandedness and fragmentation. The established protocols and the gained knowledge will be applicable for future sequencing-based high-density SNP genotyping in various fields.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: DNA Limite: Humans Idioma: En Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: DNA Limite: Humans Idioma: En Ano de publicação: 2024 Tipo de documento: Article