• Title/Summary/Keyword: SNP detection

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Improvement of SNPs detection efficient by reuse of sequences in Genotyping By Sequencing technology (유전체 서열 재사용을 이용한 Genotyping By Sequencing 기술의 단일 염기 다형성 탐지 효율 개선)

  • Baek, Jeong-Ho;Kim, Do-Wan;Kim, Junah;Lee, Tae-Ho
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.19 no.10
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    • pp.2491-2499
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    • 2015
  • Recently, the most popular technique to determine the Genotype, genetic features of individual organisms, is the GBS based on SNP from sequences determined by NGS. As analyzing the sequences by the GBS, TASSEL is the most used program to identify the genotypes. But, TASSEL has limitation that it uses only the partial sequences that is obtained by NGS. We tried to improve the efficiency in use of the sequences in order to solve the limitation. So, we constructed new data sets by quality checking, filtering the unused sequences with error rate below 0.1% and clipping the sequences considering the location of barcode and enzyme. As a result, approximately over 17% of the SNP detection efficiency was increased. In this paper, we suggest the method and the applied programs in order to detect more SNPs by using the disused sequences.

Fabrication of Nanogap-Based PNA Chips for the Electrical Detection of Single Nucleotide Polymorphism

  • Park, Dae-Keun;Park, Hyung-Ju;Lee, Cho-Yeon;Hong, Dae-Wha;Lee, Young;Choi, In-Sung S.;Yun, Wan-Soo
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.540-540
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    • 2012
  • Selective detection of single nucleotide polymorphism (SNP) of Cytochrome P450 2C19 (CYP2C19) was carried out by the PNA chips which were electrically-interfaced with interdigitated nanogap electrodes (INEs). The INEs whose average gap distance and effective gap length were about ~70 nm and ${\sim}140{\mu}m$, respectively, were prepared by the combination of the photo lithography and the surface-catalyzed chemical deposition, without using the e-beam lithography which is almost inevitable in the conventional lab-scale fabrication of the INEs. Four different types of target DNAs were successfully detected and discriminated by the INE-based PNA chips.

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Establishment of a Tm-shift Method for Detection of Cat-Derived Hookworms

  • Fu, Yeqi;Liu, Yunqiu;Abuzeid, Asmaa M.I.;Huang, Yue;Zhou, Xue;He, Long;Zhao, Qi;Li, Xiu;Liu, Jumei;Ran, Rongkun;Li, Guoqing
    • Parasites, Hosts and Diseases
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    • v.57 no.1
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    • pp.9-15
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    • 2019
  • Melting temperature shift ($T_m-shift$) is a new detection method that analyze the melting curve on real-time PCR thermocycler using SYBR Green I fluorescent dye. To establish a $T_m-shift$ method for the detection of Ancylostoma ceylanicum and A. tubaeforme in cats, specific primers, with GC tail of unequal length attached to their 5' end, were designed based on 2 SNP loci (ITS101 and ITS296) of the internal transcribed spacer 1 (ITS1) sequences. The standard curve of $T_m-shift$ was established using the standard plasmids of A. ceylanicum (AceP) and A. tubaeforme (AtuP). The $T_m-shift$ method stability, sensitivity, and accuracy were tested with reference to the standard curve, and clinical fecal samples were also examined. The results demonstrated that the 2 sets of primers based on the 2 SNPs could accurately distinguish between A. ceylanicum and A. tubaeforme. The coefficient of variation (CV) of $T_m$- values of AceP and AtuP was 0.07% and 0.06% in ITS101 and was 0.06% and 0.08% in ITS296, respectively. The minimum detectable DNA concentration was $5.22{\times}10^{-6}$ and $5.28{\times}10^{-6}ng/{\mu}l$ samples of AceP and AtuP, respectively. The accuracy of $T_m-shift$ method reached 100% based on examination of 10 hookworm DNA samples with known species. In the clinical detection of hookworm in 69 stray cat fecal sample, the $T_m-shift$ detection results were consistent with the microscopic examination and successfully differentiated between the 2-hookworm species. In conclusion, the developed method is a rapid, sensitive and accurate technique and can provide a promising tool for clinical detection and epidemiological investigation of cat-derived hookworms.

Identification of Recently Selected Mutations Driven by Artificial Selection in Hanwoo (Korean Cattle)

  • Lim, Dajeong;Gondro, Cedric;Park, Hye Sun;Cho, Yong Min;Chai, Han Ha;Seong, Hwan Hoo;Yang, Bo Suk;Hong, Seong Koo;Chang, Won Kyung;Lee, Seung Hwan
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.5
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    • pp.603-608
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    • 2013
  • Hanwoo have been subjected over the last seventy years to intensive artificial selection with the aim of improving meat production traits such as marbling and carcass weight. In this study, we performed a signature of selection analysis to identify recent positive selected regions driven by a long-term artificial selection process called a breeding program using whole genome SNP data. In order to investigate homozygous regions across the genome, we estimated iES (integrated Extended Haplotype Homozygosity SNP) for the each SNPs. As a result, we identified two highly homozygous regions that seem to be strong and/or recent positive selection. Five genes (DPH5, OLFM3, S1PR1, LRRN1 and CRBN) were included in this region. To go further in the interpretation of the observed signatures of selection, we subsequently concentrated on the annotation of differentiated genes defined according to the iES value of SNPs localized close or within them. We also described the detection of the adaptive evolution at the molecular level for the genes of interest. As a result, this analysis also led to the identification of OLFM3 as having a strong signal of selection in bovine lineage. The results of this study indicate that artificial selection which might have targeted most of these genes was mainly oriented towards improvement of meat production.

Molecular Identification of Korean Mountain Ginseng Using an Amplification Refractory Mutation System (ARMS)

  • In, Jun-Gyo;Kim, Min-Kyeoung;Lee, Ok-Ran;Kim, Yu-Jin;Lee, Beom-Soo;Kim, Se-Young;Kwon, Woo-Seang;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.34 no.1
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    • pp.41-46
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    • 2010
  • Expensive herbs such as ginseng are always a possible target for fraudulent labeling. New mountain ginseng strains have occasionally been found deep within mountain areas and commercially traded at exorbitant prices. However, until now, no scientific basis has existed to distinguish such ginseng from commonly cultivated ginseng species other than by virtue of being found within deep mountain areas. Polymerase chain reaction (PCR) analysis of the internal transcribed spacer has been shown to be an appropriate method for the identification of the most popular species (Panax ginseng) in the Panax ginseng genus. A single nucleotide polymorphism (SNP) has been identified between three newly found mountain ginseng (KGD4, KGD5, and KW1) and already established Panax species. Specific PCR primers were designed from this SNP site within the sequence data and used to detect the mountain ginseng strains via multiplex PCR. The established multiplex-PCR method for the simultaneous detection of newly found mountain ginseng strains, Korean ginseng, and foreign ginseng in a single reaction was determined to be effective. This study is the first report of scientific discrimination of "mountain ginsengs" and describes an effective method of identification for fraud prevention and for uncovering the possible presence of other, cheaper ginseng species on the market.

SNP-Based Genetic Linkage Map and Quantitative Trait Locus Mapping Associated with the Agronomically Important Traits of Hypsizygus marmoreus

  • Oh, Youn-Lee;Choi, In-Geol;Jang, Kab-Yeul;Kim, Min-Seek;Oh, Min ji;Im, Ji-Hoon
    • Mycobiology
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    • v.49 no.6
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    • pp.589-598
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    • 2021
  • White strains of Hypsizygus marmoreus are more difficult to cultivate than are brown strains; therefore, new white strain breeding strategies are required. Accordingly, we constructed the genetic map of H. marmoreus with 1996 SNP markers on 11 linkage groups (LGs) spanning 1380.49 cM. Prior to analysis, 82 backcrossed strains (HM8 lines) were generated by mating between KMCC03106-31 and the progenies of the F1 hybrid (Hami-18 × KMCC03106-93). Using HM8, the first 23 quantitative trait loci (QTLs) of yield-related traits were detected with high limit of detection (LOD) scores (1.98-9.86). The length, thickness, and hardness of the stipe were colocated on LG 1. Especially, length of stipe and thickness of stipe were highly correlated given that the correlation coefficients were negative (-0.39, p value ≤ .01). And a typical biomodal distribution was observed for lightness of the pileus and the lightness of the pileus trait belonged to the LG 8, as did traits of earliness and mycelial growth in potato dextrose agar (PDA) medium. Therefore, results for color traits can be suggested that color is controlled by a multi-gene of one locus. The yield trait was highly negatively correlated with the traits for thickness of the stipe (-0.45, p value ≤ .01). Based on additive effects, the white strain was confirmed as recessive; however, traits of mycelial growth, lightness, and quality were inherited by backcrossed HM8 lines. This new genetic map, finely mapped QTLs, and the strong selection markers could be used in molecular breeding of H. marmoreus.

Detection of superior genotype of fatty acid synthase in Korean native cattle by an environment-adjusted statistical model

  • Lee, Jea-Young;Oh, Dong-Yep;Kim, Hyun-Ji;Jang, Gab-Sue;Lee, Seung-Uk
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.6
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    • pp.765-772
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    • 2017
  • Objective: This study examines the genetic factors influencing the phenotypes (four economic traits:oleic acid [C18:1], monounsaturated fatty acids, carcass weight, and marbling score) of Hanwoo. Methods: To enhance the accuracy of the genetic analysis, the study proposes a new statistical model that excludes environmental factors. A statistically adjusted, analysis of covariance model of environmental and genetic factors was developed, and estimated environmental effects (covariate effects of age and effects of calving farms) were excluded from the model. Results: The accuracy was compared before and after adjustment. The accuracy of the best single nucleotide polymorphism (SNP) in C18:1 increased from 60.16% to 74.26%, and that of the two-factor interaction increased from 58.69% to 87.19%. Also, superior SNPs and SNP interactions were identified using the multifactor dimensionality reduction method in Table 1 to 4. Finally, high- and low-risk genotypes were compared based on their mean scores for each trait. Conclusion: The proposed method significantly improved the analysis accuracy and identified superior gene-gene interactions and genotypes for each of the four economic traits of Hanwoo.

Production of Repetitive Polypeptides for an Efficient DNA Analysis on a Microchip (Microchip상에서 효율적인 DNA 분석을 위한 반복단위 단백질의 생산)

  • Yi, Hyeon-Jin;Choi, Seok-Jin;Seo, Tae-Seok;Won, Jong-In
    • KSBB Journal
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    • v.25 no.2
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    • pp.199-204
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    • 2010
  • We generated the feasibility of DNA separation in free-solution using genetically engineered repetitive polypeptides as drag-tags. Two different-sized repetitive polypeptides were designed, expressed in E. coli, and purified. They were conjugated to a fluorescently labeled DNA (100 base), and the electrophoretic mobilities of these conjugate molecules were analyzed on a microchip. The results of these studies indicate that genetically engineered repetitive polypeptide is a prominent candidate for rapid and high-throughput genetic mutation detection, such as SNP analysis.