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Comparison of the optimal and suboptimal quantity of mitotype libraries using next-generation sequencing
ID
Obal, Marcel
(
Avtor
),
ID
Zupanc, Tomaž
(
Avtor
),
ID
Zupanič-Pajnič, Irena
(
Avtor
)
PDF - Predstavitvena datoteka,
prenos
(498,32 KB)
MD5: B1725E597F78F20AA6E82C0F49C1A300
URL - Izvorni URL, za dostop obiščite
https://link.springer.com/article/10.1007/s00414-023-03099-7
Galerija slik
Izvleček
Optimizing analysis parameters and sample input is crucial in forensic genetics methods to generate reliable results, and even more so when working with muti-copy mitochondrial DNA (mtDNA) and low-quality samples. This study compared mitotypes based on next-generation sequencing (NGS) results derived from the same samples at two different sequencing library concentrations—30 pM and 0.3 pM. Thirty femur samples from the Second World War were used as a model for poorly preserved DNA. Quantitative PCR (qPCR) method targeting 113 bp long fragment was employed to assess the quantity of mitogenomes. HID Ion Chef™ Instrument with Precision ID mtDNA Control Region Panel was used for library preparation and templating. Sequencing was performed with Ion GeneStudio™ S5 System. Reference haplotypes were determined from sequencing samples at 30 pM library input. Haplotypes were compared between optimal (30 pM) and suboptimal (0.3 pM) library inputs. Often the difference in haplotypes was length heteroplasmy, which in line with other studies shows that this type of variant is not reliable for interpretation in forensics. Excluding length variants at positions 573, 309, and 16,193, 56.7% of the samples matched, and in two samples, no sequence was obtained at suboptimal library input. The rest of the samples differed between optimal and suboptimal library input. To conclude, genotyping and analyzing low-quantity libraries derived from low-quality aged skeletonized human remains therefore must be done with caution in forensic genetics casework.
Jezik:
Angleški jezik
Ključne besede:
haplotype
,
library input
,
mitogenome
,
next-generation sequencing
,
Second World War
Vrsta gradiva:
Članek v reviji
Tipologija:
1.03 - Drugi znanstveni članki
Organizacija:
MF - Medicinska fakulteta
Status publikacije:
Objavljeno
Različica publikacije:
Objavljena publikacija
Leto izida:
2024
Št. strani:
Str. 395–400
Številčenje:
Vol. 138, iss. 2
PID:
20.500.12556/RUL-154409
UDK:
616:340.6
ISSN pri članku:
1437-1596
DOI:
10.1007/s00414-023-03099-7
COBISS.SI-ID:
167475971
Datum objave v RUL:
13.02.2024
Število ogledov:
394
Število prenosov:
37
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Objavi na:
Gradivo je del revije
Naslov:
International journal of legal medicine
Skrajšan naslov:
Int. j. leg. med.
Založnik:
Springer Nature, International Academy of Legal Medicine
ISSN:
1437-1596
COBISS.SI-ID:
512020249
Licence
Licenca:
CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
Povezava:
http://creativecommons.org/licenses/by/4.0/deed.sl
Opis:
To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.
Sekundarni jezik
Jezik:
Slovenski jezik
Ključne besede:
haplotip
,
vnos v knjižnico
,
mitogenom
,
sekvenciranje naslednje generacije
,
druga svetovna vojna
Projekti
Financer:
ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Številka projekta:
J3-3080
Naslov:
Določitev predniškega porekla iz DNA za identifikacijske namene
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