High levels of mitochondrial DNA heteroplasmy in single hair roots: Reanalysis and revision

Grzybowski, Tomasz; Malyarchuk, B. A.; Czarny, Jakub; Miścicka-Śliwka, Danuta; Kotzbach, Roman

doi:10.1002/elps.200390149

Cited by 39 publications

(17 citation statements)

References 41 publications

(54 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Various approaches and different primer sets for amplifying and sequencing the coding region and the control region (CR) (or portions thereof) have been reported [21][22][23]. It has been demonstrated that nested PCR analysis can cause an elevated number of phantom mutations [24,25], which in the case of two-stranded sequencing are manifested in pairs of conflicting basecalls at several positions. Such signals could then be misinterpreted as instances of natural heteroplasmy.…”

Section: Introductionmentioning

confidence: 98%

Phantom mutation hotspots in human mitochondrial DNA

Brandstätter¹,

Sänger²,

Lutz-Bonengel³

et al. 2005

Electrophoresis

View full text Add to dashboard Cite

Phantom mutations are systematic artifacts generated in the course of the sequencing process. Contra common belief these artificial mutations are nearly ubiquitous in sequencing results, albeit at frequencies that may vary dramatically. The amount of artifacts depends not only on the sort of automated sequencer and sequencing chemistry employed, but also on other lab-specific factors. An experimental study executed on four samples under various combinations of sequencing conditions revealed a number of phantom mutations occurring at the same sites of mitochondrial DNA (mtDNA) repeatedly. To confirm these and identify further hotspots for artifacts, > 5000 mtDNA electropherograms were screened for artificial patterns. Further, > 30 000 published hypervariable segment I sequences were compared at potential hotspots for phantom mutations, especially for variation at positions 16085 and 16197. Resequencing of several samples confirmed the artificial nature of these and other polymorphisms in the original publications. Single-strand sequencing, as typically executed in medical and anthropological studies, is thus highly vulnerable to this kind of artifacts. In particular, phantom mutation hotspots could easily lead to misidentification of somatic mutations and to misinterpretations in all kinds of clinical mtDNA studies.

show abstract

Section: Introductionmentioning

confidence: 98%

Phantom mutation hotspots in human mitochondrial DNA

Brandstätter¹,

Sänger²,

Lutz-Bonengel³

et al. 2005

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…However, it also means that maternally related individuals cannot be distinguished from one another using this technique, reducing its usefulness in criminal investigation when individualisation is required. An additional observation that has received attention in the forensic DNA community is the apparent high level of heteroplasmy that has been observed in mtDNA recovered from hairs [21][22][23][24][25][26][27]. Heteroplasmy in mtDNA sequences derived from hairs is observed as the production of two or more differing DNA sequences arising from a single individual.…”

Section: Dna Analysismentioning

confidence: 95%

DNA reviews: hair

Graham

2007

Forens Sci Med Pathol

View full text Add to dashboard Cite

Human and nonhuman hairs are often recovered during forensic investigation. As with all other biological samples that may be collected, via DNA analysis, hairs have the potential to provide the investigating authority with valuable intelligence pertaining to the identity of offenders, victims and even pets. DNA analysis of hairs is not however a straightforward process. The biochemical make up of hairs provide the DNA analyst with a unique set of challenges that must be overcome before any useful information can be gleaned from the evidence. This short review provides an overview of the location and condition of DNA within hair samples, and discusses the analytical methods that are employed to maximise the information that can be obtained from this sample type.

show abstract

“…The properties of mtDNA extracted from hair shafts have been described in depth by several authors [1,5,[7][8][9][10] and a few research articles on the possibility to use STR typing on DNA extracts have also been published in recent last years [11]. Furthermore, McNevin et al [12] recently published an optimized protocol for nuclear DNA extraction from modern hair shafts.…”

Section: Introductionmentioning

confidence: 97%