• Title/Summary/Keyword: Capillary electrophoresis

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Analysis of trinucleotide repetitive sequences for Korean patients with spinocerebellar ataxia types 8, 12, and 17

  • Kim, Gu-Hwan;Chung, Sun Ju;Ryu, Ho-Sung;Kim, Jaemin;Lee, Jin-Joo;Choi, Seoung Hoon;Lee, Juyeon;Lee, Beom Hee;Choi, Jin-Ho;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.12 no.1
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    • pp.38-43
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    • 2015
  • Purpose: Spinocerebellar ataxias (SCAs) are progressive neurodegenerative disorders with diverse modes of inheritance. There are several subtypes of SCAs. SCA 8, SCA 12, and SCA 17 are the less common forms of SCAs with limited information available on their epidemiological profiles in Korea. The purpose of this study was to investigate the prevalence of SCA8, SCA12, and SCA17 in Korea. Materials and Methods: Ninety-six unrelated Korean patients were enrolled and showed normal trinucleotide repeats through polymerase-chain reaction (PCR) for the genes ATXN1, ATXN2, ATXN3, CACNA1A, and ATXN7, which correspond to SCA1, SCA2, SCA3, SCA6, and SCA7, respectively. PCR products from patients were further analyzed by capillary electrophoresis using fluorescence labeled primers for the genes ATXN8OS, PPP2R2B, and TBP, which correspond to SCA8, SCA12, and SCA17. Results: Three patients had 104, 97, and 75 abnormal expanded repeats in the ATXN8OS gene, the causative gene for SCA8. None of the patients exhibited abnormal repeats in SCA12 and SCA17. Normal trinucleotide repeat ranges of the cohort in this study were estimated to be 17-34 copies (average, $24{\pm}4copies$) for SCA8, 7-18 copies (average, $13{\pm}3copies$) for SCA12, and 26-43 copies (average, $35{\pm}2copies$) for SCA17. Conclusion: This study demonstrated that SCA8, SCA12, and SCA17 are rare in Korean patients with SCA, and further genetic studies are warranted to enhance the mutation detection rate in the Korean SCA population.

Development of species-specific multiplex PCR assays of mitochondrial 12S rRNA and 16S rRNA for the identification of animal species (식육감별을 위한 미토콘드리아 12S rRNA와 16S rRNA 유전자의 종 특이적 multiplex PCR 기법 개발)

  • Koh, Ba-Ra-Da;Kim, Ji-Yeon;Na, Ho-Myung;Park, Seong-Do;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.34 no.4
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    • pp.417-428
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    • 2011
  • Species-specific PCR assay was developed for detection of cattle, sheep, goat, horse, dog, pig, chicken, duck, goose, and turkey using mitochondrial 12S rRNA and 16S rRNA as target genes. Also, an internal positive control was used to detect possible false negatives by using 18S rRNA gene. We designed species-specific primers with amplicon length of 190, 219, 350, 467, 241, 119, 171, 229, 111 and 268 bp for cattle, sheep, goat, horse, dog, pig, chicken, duck, goose, and turkey respectively. The specificity of the primers was tested against the other 10 non-target animal species and a cross-reaction was not observed. We developed two multiplex PCR assays for the simultaneous identification of Korea's major livestock species (cattle, pig, chicken and duck) and poultry species (chicken, duck, goose and turkey) from analogous samples, retaining the same specificity. The limit of detection of the multiplex PCR assay (cattle, pig, chicken and duck) ranged between 1 pg and 0.1 pg of template DNA extracts from raw meat. Applying multiplex PCR assays to DNA extracts from experimental pork/beef and pork/chicken tested raw and heat-treated ($120^{\circ}C$ for 30 min) mixtures respectively, detection limit was 0.1% level beef in pork, pork in beef and chicken in pork and 1.0% level pork in chicken. In conclusion, this assay using gel-based capillary electrophoresis would be very useful in highly sensitive and rapid identification of animal species or ingredients in minced meat and other meat products.

Photoalteration in Biodegradability and Chemical Compositions of Algae- derived Dissolved Organic Matter (자외선에 의한 조류기원 용존유기물의 생분해도 및 화학조성변환.)

  • Imai, Akio;Matsushige, Kazuo;Nagai, Takashi;Kim, Yong-Hwan;Kim, Bom-Chul;Choi, Kwang-Soon
    • Korean Journal of Ecology and Environment
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    • v.36 no.3 s.104
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    • pp.235-241
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    • 2003
  • The effect of ultraviolet (UV) radiation on the characteristics of algae-derived dissolved organic matter (DOM) was examined by comparing the biodegradability and DOM fraction distribution of algal DOM before and after UV exposure. Algal DOM from two axenic cultures of Microcystis aeruginosa and Oscillatoria agardhii were irradiated for 24 h at a UV intensity of 42 W/$m^2$. A complete degradation of algal DOM during the UV exposure did not occur, remaining at constant concentrations of dissolved organic carbon(DOC). After UV exposure, however, microbial degradations were reduced by 17% in M. aeruginosa and 53% in O. agardhii, respectively, and decomposition rates also were two times lower in UV exposed algal DOM. In addition, the chemical compositions of algal DOM altered substantially after UV radiation exposure. The proportions of hydrophilic bases (HiB; protein-like DOM) decreased considerably in both algal DOM sources after UV exposure (16.8% and 20.0% of DOM, respectively), whereas those of hydrophilic acids (HiA; carboxylic acids-like DOM) increased as much as the decrease of the HiB fraction. Capillary ion electrophoresis (CE) analysis showed that several carboxylic acids increased significantly after UV exposure, further confirming an increase in HiA fractions. The results of this study clearly indicate that algal DOM can be changed in its chemical composition as well as biodegradability without complete degradation by UV radiation.