• 제목/요약/키워드: pyrosequencing assay

검색결과 3건 처리시간 0.019초

Comparison of Methods for Detecting and Quantifying Variation in Copy Numbers of Duplicated Genes

  • Jeon, Jin-Tae;Ahn, Sung-Jin
    • Communications for Statistical Applications and Methods
    • /
    • 제16권6호
    • /
    • pp.1037-1046
    • /
    • 2009
  • Copy number variations(CNVs) are known as one of the most important factors in susceptibility to genetic disorders because they affect expression levels of genes. In previous studies, pyrosequencing, mini-sequencing real-time polymerase chain reaction(PCR), invader assays and other techniques have been used to detect CNVs. However, the higher the copy number in a genome, the more difficult it is to resolve the copies, so a more accurate method for measuring CNVs and assigning genotype is needed. PCR followed by a quantitative oligonucleotide ligation assay(qOLA) was developed for quantifying CNVs. The aim of this study was to compare the two methods for detecting and quantifying the CNVs of duplicated gene: the published pyrosequencing assay(pyro_CNV) and the newly developed qOLA_CNV. The accuracy and precision of the assay were evaluated for porcine KIT, which was selected as a model locus. Overall, the root mean squares(RMSs) of bias and standard deviation of qOLA_CNV were 2.09 and 0.45, respectively. These values are less than half of those of pyro CNV.

파이로시퀀싱을 이용한 한강상수원보호구역 수계 중의 세균 다양성 (Analysis of Bacterial Diversity in Water from the Han River Water Source Protection Area via a Pyrosequencing Assay)

  • 김희정;권덕인;김창수;이시원
    • 한국환경보건학회지
    • /
    • 제42권4호
    • /
    • pp.274-279
    • /
    • 2016
  • Objectives: We investigated bacterial diversity in the Han River water resource protection area in order to provide basic microbiological information on the drinking water safety of the Seoul metropolitan region. Methods: Samples were collected in the spring and winter, but not during the rainy season. Pyrosequencing, gene amplification, and extraction of nucleic acids were employed in this study. Results: In total, 57 and 48 operational taxonomic units were respectively analyzed in samples collected during spring and winter. Proteobacteria were predominant in all samples. The samples contained phylogenetically diverse bacterial communities, with eleven major phyla and 36 genera. Cyanobacteria were predominant in the spring samples, but not in the winter samples. The predominant species in the samples collected during both seasons belonged to the genus Aquamicrobium and Bradyrhizobium. Moreover, no pathogenic bacteria were detected in the samples. Conclusion: Proteobacteria were predominant in the samples from the Han River water source protection area. Cyanobacteria were more predominant in the spring samples than in the winter samples, but Aquamicrobium and Bradyrhizobium were predominant in both sampling seasons.

Quantitative Oligonucleotide Ligation Assay(qOLA)를 이용한 Landrace 품종의 KIT 유전자 반복수 변이 탐지 (Detection of Copy Number Variation of the KIT Gene in the Landrace Breed using an Quantitative Oligonucleotide Ligation Assay(qOLA))

  • 서보영;김재환;남덕우;유채경;이상호;이재봉;임현태;정은지;조인철;허강녕;전진태
    • Journal of Animal Science and Technology
    • /
    • 제49권5호
    • /
    • pp.559-568
    • /
    • 2007
  • 최근 들어 유전자 또는 DNA 분절의 반복수 변이 (CNV)에 관한 연구가 다수 수행되었으며, 그 분석 방법 및 기기 또한 다양하게 발달되었다. 본 연구는 Landrace 품종의 KIT 유전자 CNV 탐지를 위하여 다른 분석 방법들에 비하여 정확도가 높은 정량적 OLA 기법(qOLA)을 이용하였다. qOLA와 pyrosequencing assay의 조합하여 분석한 결과를 Landrace 44두에 대한 좌표 분석을 한 결과 I1/I1 또는 I3/i(IBe), I1/I2 또는 I3/IP, I1/I3, I1/IP 또는 I2/i(IBe), I2/I2, I2/IP의 6 genotype으로 분류되었으며, PROC FASTCLUS procedure을 이용한 통계 분석과 좌표 분석을 상호 비교한 결과 genotype의 분류가 100% 일치하였다. 기기간의 차이점을 조사하기 위해 qOLA_3100과 qOLA _3130의 관측치 비교를 실시한 결과 동일한 genotype 분류결과를 얻었다. 또한 정밀성 및 정확도 비교에서 qOLA_3100의 경우 표준편차와 변이계수의 평균이 2.33과 4.10로 qOLA_3130(2.67과 4.81)에 비하여 낮게 나타났다. qOLA 반응전 PCR 산물에 대하여 proteinase K 처리효과를 분석한 결과 pro- teinase K를 처리한 경우 전기영동 분석시 noise peak들이 제거되었으며 각 genotype의 이론적 비율에 보다 정확히 일치하였다.