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Development of a quantitative PCR‐based method for studying temporal DNA degradation in waterlogged bone
Author(s) -
Thornton Isis,
Cartozzo Claire,
Mays D’Arcy,
Singh Baneshwar,
Simmons Tal
Publication year - 2021
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/1556-4029.14641
Subject(s) - dna extraction , dna , genomic dna , extraction (chemistry) , taqman , real time polymerase chain reaction , polymerase chain reaction , dna profiling , digital polymerase chain reaction , biology , environmental chemistry , chromatography , chemistry , biochemistry , gene
While several studies have examined temporal DNA degradation in bones collected from terrestrial environments, studies on temporal DNA degradation in bones collected from aquatic environments are limited and mostly based on case studies. The objective of this study was to assess the impact of long‐term submersion, aquatic environment, bone type and DNA extraction method on DNA quality and quantity. Bone samples (scapulae and ribs), collected every ~1000 ADD from a freshwater lake and river, underwent DNA extraction via ChargeSwitch ® gDNA Plant Kit and organic phenol–chloroform methods, and DNA quantitation using both TaqMan and SYBR Green‐based quantitative PCR (qPCR) methods. Results suggest that in both bone types, quality of recovered DNA (i.e., degradation index) declined significantly with increase in submersion time. Among two bone types, quality of recovered DNA from scapulae declined faster than rib samples. There was no significant difference in recovered DNA quantity between bone types, DNA extraction methods, or locations but various interactions between these variables showed significant difference. Overall, it can be concluded that DNA can be extracted from waterlogged bone in sufficient quantity to generate an STR profile up to 4000 ADD.