An efficient method for the extraction of DNA from vertebrate tissues

Gemmell, Neil J.; Akiyama, Shiro

doi:10.1016/s0168-9525(96)80005-9

Cited by 256 publications

(178 citation statements)

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“…Both sample types were air dried at room temperature before being frozen (À20 1C) before analysis. Sample extraction and microsatellite analysis DNA was extracted from the dried blood and/or skin samples using a modified version of Gemmell and Akiyama's (1996) method for vertebrate tissues. Samples were placed in 300 ml of digestion solution (100 mM TRIS, 1%sodium dodecyl sulfate, 100 mM NaCl) and 7 ml of protinase K (10 mg ml À1 ), and mixed overnight (E12 h) on a rotating wheel at 37 1C.…”

Section: Methodsmentioning

confidence: 99%

Fine-scale population genetic structure and sex-biased dispersal in the smooth snake (Coronella austriaca) in southern England

et al. 2011

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Human-induced alteration of natural habitats has the potential to impact on the genetic structuring of remnant populations at multiple spatial scales. Species from higher trophic levels, such as snakes, are expected to be particularly susceptible to land-use changes. We examined fine-scale population structure and looked for evidence of sex-biased dispersal in smooth snakes (Coronella austriaca), sampled from 10 heathland localities situated within a managed coniferous forest in Dorset, United Kingdom. Despite the limited distances between heathland areas (maximum o6 km), there was a small but significant structuring of populations based on eight microsatellite loci. This followed an isolation-bydistance model using both straight line and 'biological' distances between sampling sites, suggesting C. austriaca's low vagility as the causal factor, rather than closed canopy conifer forest exerting an effect as a barrier to dispersal. Within population comparisons of male and female snakes showed evidence for sex-biased dispersal, with three of four analyses finding significantly higher dispersal in males than in females. We suggest that the fine-scale spatial genetic structuring and sex-biased dispersal have important implications for the conservation of C. austriaca, and highlight the value of heathland areas within commercial conifer plantations with regards to their future management.

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Section: Methodsmentioning

confidence: 99%

Fine-scale population genetic structure and sex-biased dispersal in the smooth snake (Coronella austriaca) in southern England

et al. 2011

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“…The actual breeding values estimated in 2006 (VIT Verden) have been assigned to the genotypes, and the associations were quantified. DNA was isolated from whole blood following the method described in Gemmel, Akiyama (1996), the quick method of KAWASAKI (1990), or from frozen sperm following the method of ASHWELL et al (1996). In cows, the proces could be potentially simplified by obtaining DNA from milk samples (BUITKAMP and GÖTZ, 2004).…”

Section: Methodsmentioning

confidence: 99%

The Breeding Values of German Holstein Sires and the <i>DGAT1</i> Polymorphism

et al. 2007

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The effect of the genotypes and alleles at nucleotide positions 10433 and 10434 of the DGAT1 gene on the breeding values for milk performance of German Holstein sires born in 1993 (n=66), and in 1998-2001 (n=200) was analyzed. The differences in breeding values had in both years the same trend. In the younger sires, the breeding value for fat percentage was higher by 0.42 % in the Lysine homozygotes compared to the Alanine homozygotes (P<0.001). A higher milk breeding value in the Alanine homozygotes (811 kg) was found compared to the Lysine homozygotes (256 kg) (P<0.001), and the fat yield breeding value was higher in the Lysine sires (DGAT1 K /DGAT1 K 29.6, DGAT1 A /DGAT1 A 15.9). The breeding value for the protein percentage was higher in the sires carrying the Lysine variant (0.02) than in the Alanine homozygotes (-0.03, P<0.05), and the protein yield was higher in the Alanine homozygotes (DGAT1 K /DGAT1 K 10.9, DGAT1 A /DGAT1 A 25.0, P<0.001). Because of the high emphasis of the index on the protein yield, the relative breeding value for milk was 110 among Alanine homozygotes, and 104 among Lysine. Comparing old and young sires, the genetic background against which the effect of DGAT1 is evaluated has changed. The selection on breeding value for protein yield increased the prevalence of DGAT1 A /DGAT1 A genotypes in protein and milk yield, and in relative breeding value for milk. In the old group, there was the highest protein yield in the heterozygous sires DGAT KA , it could be explained by another genetic background of the tested female population. Because the breeding value for fat percentage of heterozygous sires has lain within the values of homozygotes in both old and young sires, we assume the intermediary heredity on the locus. The differences of genotypic and allelic frequencies among old and young sires were not significant, however, there was certain tendency of increase of allele A (0.60 old sires, 0.64 young). On the results of this paper, the locus of DGAT1 offers information for evaluation of breed animals.

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“…Tissues were digested in (200 ml) lysis buffer with 1 ml Proteinase K (25 mg ml À1 ), and genomic DNA was purified using a lithium chloride protocol (Gemmell and Akiyama, 1996). The resulting DNA pellets were air dried and resuspended in 50 ml TE, pH 8.…”

Section: Dna Extraction and Genotypingmentioning

confidence: 99%

Genetic drift and rapid evolution of viviparity in insular fire salamanders (Salamandra salamandra)

2011

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Continental islands offer an excellent opportunity to investigate adaptive processes and to time microevolutionary changes that precede macroevolutionary events. We performed a population genetic study of the fire salamander (Salamandra salamandra), a species that displays unique intraspecific diversity of reproductive strategies, to address the microevolutionary processes leading to phenotypic and genetic differentiation of island, coastal and interior populations. We used eight microsatellite markers to estimate genetic diversity, population structure and demographic parameters in viviparous insular populations and ovoviviparous coastal and interior populations. Our results show considerable genetic differentiation (F ST range: 0.06-0.27), and no clear signs of gene flow among populations, except between the large and admixed interior populations. We find no support for island colonization by rafting or intentional/accidental anthropogenic introductions, indicating that rising sea levels were responsible for isolation of the island populations approximately 9000 years ago. Our study provides evidence of rapid genetic differentiation between island and coastal populations, and rapid evolution of viviparity driven by climatic selective pressures on island populations, geographic isolation with genetic drift, or a combination of these factors. Studies of these viviparous island populations in early stages of divergence help us better understand the microevolutionary processes involved in rapid phenotypic shifts.

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An efficient method for the extraction of DNA from vertebrate tissues

Cited by 256 publications

References 3 publications

Fine-scale population genetic structure and sex-biased dispersal in the smooth snake (Coronella austriaca) in southern England

Fine-scale population genetic structure and sex-biased dispersal in the smooth snake (Coronella austriaca) in southern England

The Breeding Values of German Holstein Sires and the <i>DGAT1</i> Polymorphism

Genetic drift and rapid evolution of viviparity in insular fire salamanders (Salamandra salamandra)

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An efficient method for the extraction of DNA from vertebrate tissues

Cited by 256 publications

References 3 publications

Fine-scale population genetic structure and sex-biased dispersal in the smooth snake (Coronella austriaca) in southern England

Fine-scale population genetic structure and sex-biased dispersal in the smooth snake (Coronella austriaca) in southern England

The Breeding Values of German Holstein Sires and the &lt;i&gt;DGAT1&lt;/i&gt; Polymorphism

Genetic drift and rapid evolution of viviparity in insular fire salamanders (Salamandra salamandra)

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The Breeding Values of German Holstein Sires and the <i>DGAT1</i> Polymorphism