A two-step procedure for extracting genomic DNA from dried blood spots on filter paper for polymerase chain reaction amplification

Zhou, Huitong; Hickford, Jonathan G. H.; Qian, Fang

doi:10.1016/j.ab.2006.03.042

Cited by 175 publications

(107 citation statements)

References 9 publications

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“…All these sheep were not closely related with the exception of NZ Romneys that were the progeny of five not closely related NZ Romney rams. Genomic DNA was purified from blood collected on FTA cards (Whatman BioScience, Middlesex, UK) according to the method described by Zhou et al (2006).…”

Section: Sheep Investigated and Dna Collectionmentioning

confidence: 99%

Variation in the ovine WFIKKN2 gene

Wang

Zhou

Qian

et al. 2014

Gene

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a b s t r a c t a r t i c l e i n f oWFIKKN2 may play a role in the regulation of muscle growth and development through its interaction with growth and differentiation factor 8 (GDF8) and growth and differentiation factor 11 (GDF11), but to date research into the function of the protein has been focused on mice, even though the WFIKKN2 gene (WFIKKN2) was first identified in humans in 2001. In this study two regions (intron 1 and the 3′ UTR) of ovine WFIKKN2 were investigated, using Polymerase Chain Reaction-Single Stranded Conformational Polymorphism (PCR-SSCP). Two different PCR-SSCP patterns, representing two unique DNA sequences (designated a and b) were detected in a 399-bp amplicon derived from the 3′ UTR, with sequence analysis revealing one single nucleotide polymorphism (SNP). In a 421-bp amplicon from intron 1, five different PCR-SSCP patterns (designated A-E) were observed and twelve SNPs were detected. Either one or two different sequences were detected in individual sheep and all the sequences identified shared homology with the WFIKKN2 sequences from cattle and other animal species, suggesting that these sequences represent variants of the ovine WFIKKN2 gene. In intron 1 of 487 sheep from eight breeds, variants B and C were the most common, followed by A, D and E. These results indicate that ovine WFIKKN2 is polymorphic and suggest that further analysis is required to see if variation in the gene is associated with variation in growth and muscle traits in sheep.

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Section: Sheep Investigated and Dna Collectionmentioning

confidence: 99%

Variation in the ovine WFIKKN2 gene

Wang

Zhou

Qian

et al. 2014

Gene

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“…Blood samples were collected onto FTA cards (Whatman BioScience, Middlesex, UK) and genomic DNA was purified using a two-step washing procedure as described in Zhou et al [9].…”

Section: Sheep Investigated and Dna Isolationmentioning

confidence: 99%

Identification of the keratin-associated protein 13-3 (KAP13-3) gene in sheep

Gong¹,

Zhou²,

Dyer³

et al. 2011

OJGen

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Keratin-associated proteins (KAPs) are a major structural component of hair and wool fibres, and play a critical role in determining the properties of the fibre. To date, forty functional high sulphur KAP genes from fourteen families have been identified in humans, but only seven functional high sulphur KAP genes have been identified in sheep. This led us to search for the ovine KAP13-3 gene, a gene encoding a high sulphur KAP. In this study, the notional KAP13-3 gene (KRTAP13-3) was amplified using primers designed based on a reported bovine KRTAP13-3 sequence. PCR-single stranded conformational polymorphism (PCR-SSCP) analysis was used to screen amplicons derived from the gene in one hundred and forty seven New Zealand Romney cross-bred sheep. Five unique banding patterns were revealed. Either one PCR-SSCP pattern (homozygous) or a combination of two patterns (heterozygous) was observed for each sheep. Sequencing of PCR amplicons representtative of different SSCP patterns revealed five different DNA sequences. The sequences derived from the amplicons showed a low homology to other known ovine KRTAPs, but had a high homology with previous reported KRTAP13-n sequences from human and cattle, with the closest homology being with bovine KRTAP13-3, suggesting the sequences represent the ovine KRTAP13-3 locus. Among the five allele sequences, four nucleotide substitutions were identified within the coding region. Of these substitutions, three were non-synonymous and would result in amino acid changes (p.Arg79Cys, p.Arg81Gln and p.Tyr130His). This variation in the KAP13-3 gene may affect gene expression, the structure and assembly of the protein, and consequently influence wool traits, if KAP13-3 is of importance to wool fibre structure.

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“…For each sample, a single FTA card punch was placed in a 200-μL tube and the genomic DNA purifi ed using a 2-step washing procedure as described by Zhou et al (2006). The ovine IGF1R gene was genotyped using PCR-single strand conformational polymorphism (PCR-SSCP) as described previously (Byun et al, 2008).…”

Section: Genotype Of Ovine Igf1rmentioning

confidence: 99%

An association between lifespan and variation in insulin-like growth factor I receptor in sheep1

Byun

Forrest

Frampton

et al. 2012

Journal of Animal Science

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Longevity in livestock is a valuable trait. When productive animals live longer, fewer replacement animals need to be raised. However, selection for longevity is not commonly the focus of breeding programs as direct selection for long-lived breeding stock is virtually impossible until late in the reproductive life of the animal. Additionally the underlying genetic factors or genes associated with longevity are either not known, or not well understood. In humans, there is evidence that IGF 1 receptor (IGF1R) is involved in longevity. Polymorphism in the IGF1R gene has been associated with longevity in a number of species. Recently, 3 alleles of ovine IGF1R were identifi ed, but no analysis of the effect of IGF1R variation on sheep longevity has been reported. In this study, associations between ovine IGF1R variation, longevity and fertility were investigated.Polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) was used to type IGF1R variation in 1,716 New Zealand sheep belonging to 6 breeds and 36 fl ocks. Ovine IGF1R C was associated with age when adjusting for fl ock (present 5.5 ± 0.2 yr, absent 5.0 ± 0.1 yr, P = 0.02). A general linear mixed effects model suggested an association (P = 0.06) between age and genotype, when correcting for fl ock. Pairwise comparison (least signifi cant difference) of specifi c genotypes revealed the difference to be between AA (5.0 ± 0.1 yr) and AC (5.6 ± 0.2 yr, P = 0.02). A weak negative Pearson correlation between fertility and longevity traits was observed (r = -0.25, P < 0.01). The fi nding of an association between variation in IGF1R and lifespan in sheep may be useful in prolonging the lifespan of sheep.

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A two-step procedure for extracting genomic DNA from dried blood spots on filter paper for polymerase chain reaction amplification

Cited by 175 publications

References 9 publications

Variation in the ovine WFIKKN2 gene

Variation in the ovine WFIKKN2 gene

Identification of the keratin-associated protein 13-3 (KAP13-3) gene in sheep

An association between lifespan and variation in insulin-like growth factor I receptor in sheep1

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