Comparative Identity and Homogeneity Testing of the mtDNA HV1 Region Using Denaturing Gradient Gel Electrophoresis

Steighner, Robert J.; Tully, Lois A.; Karjala, Justin D.; Coble, Mike D.; Holland, Mitchell M.

doi:10.1520/jfs14586j

Cited by 14 publications

(11 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the DGGE assay, in a comparison of the samples, when the PCR products of each sample are identical, each band showed the same mobility on the gel and when the PCR products are not identical, each band could show a different mobility on the gel. If heteroplasmic positions were present in PCR products that contained two different sequences, two homoduplex bands and two heteroduplex bands were normally observed in the gel (22). Most of the 480 hair samples showed a single homoduplex band with the same mobility as the blood samples, indicating no heteroplasmy, but 28 samples among three individuals and three regions (DGGE-B, DGGE-C and DGGE-E) showed bands that were different from the blood samples, indicating either a mutation or heteroplasmy.…”

Section: Resultsmentioning

confidence: 99%

“…After the electrophoresis, the gels were stained with 1 × SYBR Green I (Molecular Probes, Eugene, OR) for 20 min and visualized using Fluorimager 595 (Amersham-Pharmacia Biotech). The relative proportion of each band in the heteroplasmic samples was determined by calculating the peak intensities from the scanned gel image using the NIH image software (developed at the United States National Institutes of Health) (22).…”

Section: Denaturing Gradient Gel Electrophoresis (Dgge)mentioning

confidence: 99%

“…Because of the high level of heteroplasmy, the latter report has been questioned and can be regarded as controversial (18,19) heteroplasmy and several mutations in hairs from a single individual during an interlaboratory study (21). A denaturing gradient-gel electrophoresis (DGGE) assay has been demonstrated to be a sensitive tool for detecting heteroplasmy (6,22). The use of a DGGE assay for the detection of mutations or heteroplasmic positions in the HV1 region has been reported (22).…”

Section: Introductionmentioning

confidence: 99%

“…A denaturing gradient-gel electrophoresis (DGGE) assay has been demonstrated to be a sensitive tool for detecting heteroplasmy (6,22). The use of a DGGE assay for the detection of mutations or heteroplasmic positions in the HV1 region has been reported (22). The detection limit was investigated and the DGGE assay was found to be more sensitive in the detection of heteroplasmy than sequencing analysis (6).…”

mentioning

confidence: 99%

See 3 more Smart Citations

Mitochondrial DNA Heteroplasmy Among Hairs from Single Individuals

Sekiguchi

Sato

Kasai

2004

Journal of Forensic Sciences

View full text Add to dashboard Cite

A denaturing gradient gel electrophoresis (DGGE) assay was used to detect mitochondrial DNA (mtDNA) sequence heteroplasmy in 160 hairs from each of three individuals. The HV1 and HV2 heteroplasmic positions were then identified by sequencing. In several hairs, the heteroplasmic position was not evident by sequencing and dHPLC separation of the homoduplex/heteroduplex species was carried out with subsequent reamplification and sequencing to identify the site. The overall detection frequency of sequence heteroplasmy in these hairs was 5.8% (28/480) with DGGE and 4.4% (21/280) with sequencing. Sequence heteroplasmy of hair was observed even when the reference blood sample of the individual was homoplasmic. The heteroplasmic positions were not necessarily observed at sites where high rates of substitution have been reported. In two hairs, a complete single base change from the reference blood sample was observed with sequencing, while the heteroplasmic condition at that site in the hair was observed using DGGE. The DGGE results in such samples would serve as an aid in considering the possibility of match significance. In a forensic case, this situation would lead to the possibility of a failure to exclude rather than to be inconclusive. KEYWORDS:forensic science, DNA typing, mitochondrial DNA, hair DNA, heteroplasmy, denaturing gradient gel electrophoresis MtDNA analysis is a useful tool for the identification of old or degraded biological evidentiary samples and is also important in the forensic comparison of hair samples. Heteroplasmy, the presence of two or more types of mtDNA in a single individual, was first observed in humans in association with a mitochondrial disorder (1). However, heteroplasmy in mtDNA has been reported, not only in association with mitochondrial disorders, but in the non-coding control region as well. Heteroplasmy in the control region was initially not considered to be significant (2), but it now appears to be a more frequent phenomenon than had originally been thought (3-6). Inter-and intra-generational heteroplasmy has been reported and the proportion of heteroplasmy appears to vary among different family members (5,7,8). Intra-individual heteroplasmy has also been reported in several studies (9,10). Length heteroplasmy in the HV1 and HV2 region, near nucleotide positions 16189 and 309-315, was found among hairs within a single individual (11,12) and the mechanism involved in this length heteroplasmy has been investigated (13,14). Site heteroplasmy, at a single nucleotide position in the control region of mtDNA in hairs, has also been reported and its proportions varied among hairs within a single individual (5,15). The frequency of site heteroplasmy among hairs from an individual with a homoplasmic blood sample has also been reported (16,17). Hühne et al. reported that no heteroplasmy was detected in an analysis of 15 hairs from each of 10 individuals, whereas Grzybowski reported numerous heteroplasmic positions in an analysis of 2-6 hairs each from each of 13 individuals. Be...

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Denaturing Gradient Gel Electrophoresis (Dgge)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Mitochondrial DNA Heteroplasmy Among Hairs from Single Individuals

Sekiguchi

Sato

Kasai

2004

Journal of Forensic Sciences

View full text Add to dashboard Cite

show abstract

“…Many have shown superior sensitivity over sequencing, which typically requires that the minor heteroplasmic sequence be present at u minimum concentration of lO-200/0 (Bendall et a/., 1996). For example, a DGGE assay designed to examine heteroplasmy in hypervariable region I (HV 1) demonstrated sufficient sensitivity to detect a heteroplasmic species present at concentrations as low as I% (Tully, 1998;Steighner et al, 1999). Detection and quantification of the 11,778 Inutation (LHON) at levels as low as 1.50/0 has been achieved using DNA minisequencing (Juvonen et al, 1994).…”

Section: ;mentioning

confidence: 99%

Human Mitochondrial Genetics

Tully

Levin²

2000

Biotechnology and Genetic Engineering Reviews

Self Cite

View full text Add to dashboard Cite

Terrestrial nest‐building by wild chimpanzees (Pan troglodytes): Implications for the tree‐to‐ground sleep transition in early hominins

Koops

McGrew

Matsuzawa

et al. 2012

American J Phys Anthropol

View full text Add to dashboard Cite

Nest-building is a great ape universal and arboreal nesting in chimpanzees and bonobos suggests that the common ancestor of Pan and Homo also nested in trees. It has been proposed that arboreal nest-building remained the prevailing pattern until Homo erectus, a fully terrestrial biped, emerged. We investigated the unusual occurrence of ground-nesting in chimpanzees (Pan troglodytes), which may inform on factors influencing the tree-to-ground sleep transition in the hominin lineage. We used a novel genetic approach to examine ground-nesting in unhabituated chimpanzees at Seringbara in the Nimba Mountains, Guinea. Previous research showed that ground-nesting at Seringbara was not ecologically determined. Here, we tested a possible mate-guarding function of ground-nesting by analyzing DNA from shed hairs collected from ground nests and tree nests found in close proximity. We examined whether or not ground-nesting was a group-level behavioral pattern and whether or not it occurred in more than one community. We used multiple genetic markers to identify sex and to examine variation in mitochondrial DNA control region (HV1, HV2) sequences. Ground-nesting was a male-biased behavior and males constructed more elaborate ("night") nests than simple ("day") nests on the ground. The mate-guarding hypothesis was not supported, as ground and associated tree nests were built either by maternally-related males or possibly by the same individuals. Ground-nesting was widespread and likely habitual in two communities. We suggest that terrestrial nest-building may have already occurred in arboreally-adapted early hominins before the emergence of H. erectus.

show abstract

Comparative Identity and Homogeneity Testing of the mtDNA HV1 Region Using Denaturing Gradient Gel Electrophoresis

Cited by 14 publications

References 60 publications

Mitochondrial DNA Heteroplasmy Among Hairs from Single Individuals

Mitochondrial DNA Heteroplasmy Among Hairs from Single Individuals

Human Mitochondrial Genetics

Terrestrial nest‐building by wild chimpanzees (Pan troglodytes): Implications for the tree‐to‐ground sleep transition in early hominins

Contact Info

Product

Resources

About