Skip to main content
main-content

29.05.2018 | Short Communication | Ausgabe 3/2019

International Journal of Legal Medicine 3/2019

The effects of extra PCR cycles when amplifying skeletal samples with the GlobalFiler® PCR Amplification Kit

Zeitschrift:
International Journal of Legal Medicine > Ausgabe 3/2019
Autoren:
Michelle Harrel, David Gangitano, Sheree Hughes-Stamm
Wichtige Hinweise

Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1007/​s00414-018-1860-2) contains supplementary material, which is available to authorized users.

Abstract

When samples with low amounts of DNA are amplified using short tandem repeats (STRs), stochastic effects such as allele and locus dropout or drop-in, allele imbalance, and increased stutter often occur making data interpretation more difficult. The most common approach to improving STR results from low template samples is to increase the number of PCR cycles. Although more alleles may be recovered, stochastic effects may be exaggerated resulting in more complicated STR profiles. This work reports the effect of additional PCR cycles (29 vs. 30, 31, and 32) on STR success from environmentally challenged bone and tooth samples using the GlobalFiler® DNA Amplification Kit (Thermo Fisher Scientific). In addition, we compared the efficiency of two DNA extraction kits for skeletal samples: QIAamp® DNA Investigator (QIAGEN) and PrepFiler® BTA™ Forensic DNA Extraction (Thermo Fisher Scientific) kits. Results showed that more DNA was recovered from samples using the PrepFiler® BTA™ kit; but regardless of the extraction method, the number of alleles detected and the peak heights both increased with an increase in PCR cycle number. Although more alleles were reported in almost all samples, the most notable improvement was observed in samples with the DNA template < 120 pg. A general increase in the number of PCR artifacts was detected in STR profiles generated using 30–32 cycles. Overall, this study provides supporting evidence that STR profile completeness and quality may be improved when low template skeletal samples are amplified with extra PCR cycles (up to 32 cycles) using the GlobalFiler® DNA Amplification Kit.

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

★ PREMIUM-INHALT
e.Med Interdisziplinär

Mit e.Med Interdisziplinär erhalten Sie Zugang zu allen CME-Fortbildungen und Fachzeitschriften auf SpringerMedizin.de. Zusätzlich können Sie eine Zeitschrift Ihrer Wahl in gedruckter Form beziehen – ohne Aufpreis.

Zusatzmaterial
ESM 1 (DOCX 20 kb)
414_2018_1860_MOESM1_ESM.docx
ESM 2 (DOCX 1208 kb)
414_2018_1860_MOESM2_ESM.docx
ESM 3 (DOCX 142 kb)
414_2018_1860_MOESM3_ESM.docx
ESM 4 (DOCX 36 kb)
414_2018_1860_MOESM4_ESM.docx
ESM 5 (DOCX 121 kb)
414_2018_1860_MOESM5_ESM.docx
Literatur
Über diesen Artikel

Weitere Artikel der Ausgabe 3/2019

International Journal of Legal Medicine 3/2019 Zur Ausgabe

Neu im Fachgebiet Rechtsmedizin

14.03.2019 | Originalien | Ausgabe 2/2019

Medikamentennachweise bei bayerischen Altenheimbewohnern – eine rechtsmedizinische Analyse

Erste Ergebnisse aus Screeninguntersuchungen an Urinproben

07.03.2019 | Mammakarzinom | CME | Ausgabe 2/2019

Update der S3-Leitlinie Mammakarzinom

Was gibt es Neues für Pathologen?

21.02.2019 | Schwerpunkt: Neuropathologie | Ausgabe 2/2019

Neuropathologie der Medulloblastome und anderer embryonaler Tumoren des ZNS

Präzisierung der Diagnostik durch Integration genetischer Marker