RT Journal Article
T1 Adapting an established Ampliseq microhaplotype panel to nanopore sequencing through direct PCR
A1 Casanova Adán, Lucía
A1 Mosquera Miguel, Ana
A1 González Bao, Javier
A1 Ambroa Conde, Adrián
A1 Ruiz Ramírez, Jorge
A1 Cabrejas Olalla, Amaia
A1 González Martín, E.
A1 Freire Aradas, Ana María
A1 Rodríguez López, Amelia
A1 Phillips, Christopher Paul
A1 Lareu Huidobro, María Victoria
A1 Puente Vila, María del Carmen de la
K1 Nanopore sequencing
K1 MinION
K1 Microhaplotypes
K1 Kinship testing
K1 Direct PCR
K1 Field laboratories
AB We have adapted an established Ampliseq microhaplotype panel for nanopore sequencing with the Oxford Nanopore Technologies (ONT) system, as a cost-effective and highly scalable solution for forensic genetics applications. For this purpose, we designed a protocol combining direct PCR amplification from unextracted DNA with ONT library construction and sequencing using the MinION device and workflow. The analysis of reference samples at input amounts of 5–10 ng of DNA demonstrates stable coverage patterns, allele balance, and strand bias, reaching profile completeness and concordance rates of ∼95%. Similar levels were achieved when using direct-PCR from blood, buccal and semen swabs. Dilution series results indicate sensitivity is maintained down to 250 pg of input DNA, and informative profiles are produced down to 62.5 pg. Finally, we demonstrated the forensic utility of the nanopore workflow by analyzing two third degree pedigrees that showed low likelihood ratio values after the analysis of an extended panel of 38 STRs, achieving likelihood ratios 2–3 orders of magnitude higher when testing with the MinION–based haplotype data
PB Elsevier
SN 1872-4973
YR 2023
FD 2023
LK http://hdl.handle.net/10347/33244
UL http://hdl.handle.net/10347/33244
LA eng
NO Forensic Science International: Genetics, Volume 67, 2023, 102937
NO This work was partially supported by the Consellería de Cultura, Educación e Ordenación Universitaria e da Consellería de Economía, Emprego e Industria from Xunta de Galicia, Spain (ED481D-2021–008). JRR is supported by the “Programa de axudas á etapa predoutoral” funded by the Consellería de Cultura, Educación e Ordenación Universitaria e da Consellería de Economía, Emprego e Industria from Xunta de Galicia, Spain (ED481A-2020/039). MVL is supported by the Ministerio de Educación, Cultura y Ciencia, Spain (PID2019–107876RB-I00). MdlP is supported by a postdoctoral fellowship awarded by the Gobierno de España: IJC2020–042638-I, funded by MCIN/AEI/10.13039/501100011033 and the European Union "NextGenerationEU/PRTR"
DS Minerva
RD 28 abr 2026