Development and optimization of the VISAGE basic prototype tool for forensic age estimation.
Aging
/ genetics
CpG Islands
DNA Methylation
Fatty Acid Elongases
/ genetics
Forensic Genetics
/ methods
Genetic Markers
High-Throughput Nucleotide Sequencing
Humans
Intracellular Signaling Peptides and Proteins
/ genetics
Kruppel-Like Transcription Factors
/ genetics
LIM-Homeodomain Proteins
/ genetics
Multiplex Polymerase Chain Reaction
Muscle Proteins
/ genetics
Reproducibility of Results
Transcription Factors
/ genetics
Tripartite Motif Proteins
/ genetics
Age estimation
Bisulfite PCR multiplex development
MiSeq FGx sequencing
Targeted bisulfite sequencing
Journal
Forensic science international. Genetics
ISSN: 1878-0326
Titre abrégé: Forensic Sci Int Genet
Pays: Netherlands
ID NLM: 101317016
Informations de publication
Date de publication:
09 2020
09 2020
Historique:
received:
29
11
2019
revised:
22
04
2020
accepted:
03
06
2020
pubmed:
24
6
2020
medline:
13
7
2021
entrez:
24
6
2020
Statut:
ppublish
Résumé
The VISAGE (VISible Attributes through GEnomics) consortium aims to develop, optimize and validate prototype tools to broaden the use of DNA intelligence methods in forensic routine laboratories. This includes age estimation based on the quantification of DNA methylation at specific CpG sites. Here, we present the VISAGE basic prototype tool for age estimation targeting 32 CpGs from five genes ELOVL2, MIR29B2CHG (herein, MIR29B2C), FHL2, TRIM59 and KLF14. The assay interrogates these well described age markers by multiplex PCR for bisulfite converted DNA and massively parallel sequencing on a MiSeq FGx instrument. We describe protocol optimizations including tests on five bisulfite conversion kits and an evaluation of the assay's reproducibility and sensitivity with artificially methylated DNA standards. We observed robust quantification of methylation levels with a mean standard deviation of 1.4 % across ratios. Sensitivity tests showed no increase of variability down to 20 ng DNA input into bisulfite conversion with a median difference below 1.6 % between technical replicates.
Identifiants
pubmed: 32574993
pii: S1872-4973(20)30095-8
doi: 10.1016/j.fsigen.2020.102322
pii:
doi:
Substances chimiques
ELOVL2 protein, human
0
FHL2 protein, human
0
Genetic Markers
0
Intracellular Signaling Peptides and Proteins
0
KLF14 protein, human
0
Kruppel-Like Transcription Factors
0
LIM-Homeodomain Proteins
0
Muscle Proteins
0
TRIM59 protein, human
0
Transcription Factors
0
Tripartite Motif Proteins
0
Fatty Acid Elongases
EC 2.3.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
102322Informations de copyright
Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.