• Asian-Australasian Journal of Animal Sciences
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• v.15 no.9
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• pp.1237-1243
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• 2002

We have developed a reliable and noninvasive method for swine genotyping of single locus nuclear gene with aged single hair follicles delivered by general mail. The method is based on booster and nested PCR amplification with step-wise increase of primers and dNTPs concentrations followed by restriction endonuclease digestion. To establish this method, the ryanodine receptor (RYR 1) locus which is an economically important trait in swine industry was employed for genotyping experiment. The 3-step PCR amplication method is much less dependent on the quantity and quality of template DNA and produces enough amplification product for the detection on the ethidium bromide-stained gel such as RFLP analysis. A total of 120 pigs were subjected to the RYR 1 genotyping analysis using three-step PCR method which amplified enough quantity of PCR products from the aged single hair follicles for RFLP analysis and genotyping results were identical to the results of the corresponding ethanol-fixed skeletal muscle tissue. This approach will be a great help for porcine breeders and investigators in genotyping of swine. They can receive genotyping results later by simply plucking single hairs of their pigs at farm and sending them in general mail to the diagnostic laboratory which eliminates the inconveniences to collect ear tissue or blood cells from pigs, or the investigator's need for travel to farms in order to collect fresh hair samples.

• Plant Breeding and Biotechnology
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• v.6 no.4
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• pp.434-443
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• 2018

Genotyping-by-sequencing (GBS) has been used as a viable single nucleotide polymorphism (SNP) validation method that provides reduced representation sequencing by using restriction endonucleases. Although GBS makes it possible to perform marker discovery and genotyping simultaneously with reasonable costs and a simple molecular biology workflow, the standard TASSEL-GBS pipeline was designed for homozygous groups, and genotyping of heterozygous groups is more complicated. To addresses this problem, we developed a GBS pipeline for heterozygous groups that called KNU-GBS pipeline, specifically for apple (Malus domestica). Using KNU-GBS pipeline, we constructed a genetic linkage map consisting of 1,053 SNP markers distributed over 17 linkage groups encompassing a total of 1350.1 cM. The novel GBS pipeline for heterozygous groups will be useful for marker-assisted breeding programs, and diverse heterozygous genome analyses.

• Genomics & Informatics
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• v.6 no.1
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• pp.54-58
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• 2008

Genotyping Tool for Viral SEQuences (GTVseq) provides scientists with the genotype information on the viral genome sequences including HIV-1, HIV-2, HBV, HCV, HTLV-1, HTLV-2, poliovirus, enterovirus, flavivirus, Hantavirus, and rotavirus. GTVseq produces alternative and additive genotype information for the query viral sequences based on two different, but related, scoring methods. The genotype information produced is reported in a graphical manner for the reference genotype matches and each graphical output is linked to the detailed sequence alignments between the query and the matched reference sequences. GTVseq also reports the potential 'repeats' and/or 'recombination' sequence region in a separated window. GTVseq does not replace completely other well-known genotyping tools such as NCBI's virus sequence genotyping tool (http://www.ncbi. nlm.nih.gov/projects/genotyping/formpage.cgi), but provides additional information useful in the confirmation or for further investigation of the genotype(s) for the newly isolated viral sequences.

• Korean Journal of Clinical Laboratory Science
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• v.36 no.1
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• pp.1-6
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• 2004

The enterobacterial repetitive intergenic consensus (ERIC)-PCR is a recently described DNA fingerprinting technique based on amplification of repetitive element distributed in bacteria. We applied of ERIC-PCR to clinical isolates of Vibrio parahaemolyticus and other bacteria associated diarrhea. Twenty isolates of V. parahaemolyticus were used for intragenic genotyping, which were isolated from 2001 to 2002 in Chungbuk National University hospital. For interspecies genotyping, V. vulnificus, V. alginolyticus, V. parahaemolyticus, Escherichia coli, Salmonella and Shigella spp. were used. The genotyping were analyzed by ERIC-PCR. The genotyping of V. parahaemolyticus were grouped two major pattern (A, B) and were subdivided into 10 subtypes (A1, A2, B1-B8) by ERIC-PCR. These method distinctly differentiated bacterial species associated diarrhea. Those results show that ERIC-PCR can be reliable and efficient method for genotyping of V. parahaemolyticus and bacteria associated diarrhea.

• Genomics & Informatics
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• v.3 no.1
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• pp.1-7
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• 2005

Case-control studies are widely used for disease gene mapping using individual genotyping data. However, analyses of large samples are often impractical due to the expense of individual genotyping. The use of DNA pooling can significantly reduce the number of genotyping reactions required; hence reducing the cost of large-scale case-control association studies. Here, we discuss the design and analysis of DNA pooling genetic association studies.

• Proceedings of the Korean Information Science Society Conference
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• 2004.04b
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• pp.586-588
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• 2004

최근 DNA 관련 기술의 개발이 활발하게 이루어지고 있고, 그에 따라 DNA를 개인인식에 사용마고자 하는 시도가 실시간 이용가능성이 높은 DNA 칩 기술과 합쳐져 매우 중요한 이슈로 떠오르고 있다. DNA를 분석하기 위해서는 생물학적 실험이 필수적으로 따르게 되는데 이러한 실험 결과를 인식에 적용하기 위해서는 적절한 전처리가 필요하다. 본 논문에서는 여러 장점들로 인해 최근 DNA분석 기술로 주목받고 있는 모세관 전기영동법을 사용하여 DNA를 분석하고, 그 분석물을 개인인식을 위해 genotyping하는 과정에서 전처리가 요구되는 각 경우들에 대해 논하고 적절한 필터링 기법들을 제시한다.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.58-58
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• 2017

The advent of next generation sequencing technology has elicited plenty of sequencing data available in agriculturally relevant plant species. For most crop species, it is too expensive to obtain the whole genome sequence data with sufficient coverage. Thus, many approaches have been developed to bring down the cost of NGS. Genotyping-by-sequencing (GBS) is a cost-effective genotyping method for complex genetic populations. GBS can be used for the analysis of genomic selection (GS), genome-wide association study (GWAS) and constructing haplotype and genetic linkage maps in a variety of plant species. For efficiently dealing with plant GBS data, the TASSEL-GBS pipeline is one of the most popular choices for many researchers. TASSEL-GBS is JAVA based a software package to obtain genotyping data from raw GBS sequences. Here, we describe application of GBS and bioinformatics pipeline of TASSEL-GBS for analyzing plant genetics data.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.5
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• pp.361-367
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• 2005

Single nucleotide polymorphisms (SNPs) are valuable DNA markers due to their abundance and potential for use in automated high-throughput genotyping. Numerous SNP genotyping assays have been developed. In this report, one of effective and high throughput SNP genotyping assays, which was named the template-directed dye-terminator incorporation with fluorescence polarization detection (FP-TDI) was described. Although the most of this assay succeed, the objective of this work was to deter­mine the reasons for the failures, find ways to improve the assay and reduce the running cost. Ninety $F_2$-derived soybean, Glycine max (L.) Merr., RILs from a cross between 'Pureunkong' and 'Jinpumkong 2' were genotyped at four SNPs. FP measurement was done on $Victot^3$ microplate reader (perkinelmer Inc., Boston, MA, USA). Increasing the number of thermal cycles in the single-base extension step increased the separation of the FP values between the products corresponding to different genotypes. But in some assays, excess of heterozygous genotypes was observed with increase of PCR cycles. We discovered that the excess heterozygous was due to misincorporation of one of the dye­terminators during the primer extension reaction. After pyrophosphatase incubation and thermal cycle control, misincoporation can be effectively prevented. Using long amplicons instead of short amplicons for SNP genotyping and decreasing the amount of dye terminator and Acyclopol Taq polymerase to 1/2 or 1/3 decreased the cost of the assay. With these minor adjustments, the FP-TDI assay can be used more accurately and cost-effectively.

• Journal of the korean academy of Pediatric Dentistry
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• v.24 no.1
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• pp.65-81
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• 1997

For the purpose of evaluating the appropriateness of AP-PCR as a facile, rapid and reproducible method for genotyping Streptococcus mutans, and selecting the discriminant primer for it, a DNA fingerprinting was performed on the microorganisms isolated from caries-free children and children with rampant caries, respectively. In the course of selecting appropriate primer for S. mutans genotyping, we chose S2 primer from 6 different primers which shows highest resolution on the agarose gel as well. Nineteen kinds of fingerprint patterns were observed in caries-free children and children with rampant caries which were produced by combination of 7 different fragments. Interestingly, the number of types observed in caries-free children was greater than that in children with rampant caries. And we observed Type 2 was predominant in children with rampant caries (about 80%) and relatively even distribution of each types in caries-free children. Furthermore, it was appeared that the major types in normal control were not or rarely found in children with rampant caries. In conclusion, we could establish simple, rapid and highly reproducible AP-PCR method for genotyping S. mutans. We also found differences in distribution of S. mutans between normal and patient, which suggested that cariogenicity is also dependent on qualitative aspects which is caused by the difference in genotypes of S. mutans in oral cavity.

• BMB Reports
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• v.46 no.6
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• pp.305-309
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• 2013

The determination of relatedness between individuals in a family is crucial in analysis of common complex diseases. We present a method to infer close inter-familial relationships based on SNP genotyping data and provide the relationship coefficient of kinship in Korean families. We obtained blood samples from 43 Korean individuals in two families. SNP data was obtained using the Affymetrix Genome-wide Human SNP array 6.0 and the Illumina Human 1M-Duo chip. To measure the kinship coefficient with the SNP genotyping data, we considered all possible pairs of individuals in each family. The genetic distance between two individuals in a pair was determined using the allele sharing distance method. The results show that genetic distance is proportional to the kinship coefficient and that a close degree of kinship can be confirmed with SNP genotyping data. This study represents the first attempt to identify the genetic distance between very closely related individuals.