• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.372-378
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• 2017

For comparison of the transcription profiles in apple (Malus domestica L.) cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red flesh apple cultivar, 'Redfield' and white flesh apple cultivar, 'Granny Smith' were investigated by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the red flesh apple cultivar and white flesh apple cultivar were selected for nucleotide sequence determination and homology searches. High resolution melting (HRM) technique measures temperature induced strand separation of short PCR amplicons, and is able to detect variation as small as one base difference between red flesh apple cultivars and white flesh apple cultivars. We applied high resolution melting (HRM) analysis to discover single nucleotide polymorphisms (SNP) based on the predicted SNP information derived from the apple EST database. All 103 pairs of SNPs were discriminated, and the HRM profiles of amplicons were established. Putative SNPs were screened from the apple EST contigs by HRM analysis displayed specific difference between 10 red flesh apple cultivars and 11 white flesh apple cultivars. In this study, we report an efficient method to develop SNP markers from an EST database with HRM analysis in apple. These SNP markers could be useful for apple marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in apple cultivars.

• Korean Journal of Environmental Biology
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• v.36 no.2
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• pp.165-173
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• 2018

We sampled vomit of black-faced spoonbills(Platalea minor) during the brood-rearing season (from June 2011 to June 2014) at the Namdong reservoir in Incheon and analyzed the food components in the vomit using microscopy and next-generation sequencing (NGS). Microscopic observations primarily helped in identifying osteichthyes (bony fishes), crustaceans, and polychaetes. In particular, species belonging to the families Mugilidae and Gobiidae among the fish, and Macrophthalmus japonicas among the crustaceans, were observed at high frequency. Results of NGS analysis revealed the predominant presence of bony fish (42.58% of total reads) and crustaceans (40.75% of total reads), whereas others, such as polychaetes (12.66%), insects (0.24%), and unidentified species (3.78%), occurred in lower proportions. At the species level, results of NGS analysis revealed that Macrophthalmus abbreviates and Macrobrachium sp. among the crustaceans, and Acanthogobius hasta, Tridentiger obscurus, and Pterogobius zacalles among the bony fish, made up a high proportion of the total reads. These food species are frequently found at tidal flats in the Songdo and Sihwa lakes, emphasizing the importance of these areas as potential feeding sites of the black-faced spoonbill. Feed composition of the black-faced spoonbill, as evaluated by analyzing its vomit, differed when the evaluations were done by microscopic observation or by NGS analysis. Evaluation by microscopic observation is difficult and not error free, owing to the degradation of the samples to be analyzed; however, NGS analysis is more accurate, because it makes use of genetic information. Therefore, accurately analyzing food components from morphologically indistinguishable samples is possible by using genetic analysis.

• The Korean Journal of Blood Transfusion
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• v.29 no.3
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• pp.310-319
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• 2018

Background: Research on next-generation sequencing (NGS)-based HLA typing is active. To resolve the phase ambiguity and long turn-around-time of conventional high resolution HLA typing, this study developed a NGS-based high resolution HLA typing method that can handle large-scale samples within an efficient testing time. Methods: For HLA NGS, the condition of nucleic acid extraction, library construction, PCR mechanism, and HLA typing with bioinformatics were developed. To confirm the accuracy of the NGS-based HLA typing method, the results of 192 samples HLA typed by SSOP and 28 samples typed by SBT compared to NGS-based HLA-A, -B and -DR typing. Results: DNA library construction through two-step PCR, NGS sequencing with MiSeq (Illumina Inc., San Diego, USA), and the data analysis platform were established. NGS-based HLA typing results were compatible with known HLA types from 220 blood samples. Conclusion: The NSG-based HLA typing method could handle large volume samples with high-throughput. Therefore, it would be useful for HLA typing of bone marrow donation volunteers.

• Journal of Life Science
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• v.33 no.1
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• pp.25-33
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• 2023

The fermentation process of kimchi, which is a traditional Korean food, influences the resulting compo- sition of microorganisms, such as the genera Leuconostoc, Weissella, and Lactobacillus. In addition, several factors, including the type of kimchi, fermentation conditions, materials, and ingredients, can influence the distribution of the kimchi microbial community. In this study, next-generation sequencing (NGS) of kimchi samples obtained from central (Gangwon-do and Gyeonggi-do) and southern (Jeolla-do and Gyeongsang-do) regions in Korea was performed, and the microbial communities in samples from the two regions were compared. Good's coverage prediction for all samples was higher than 99%, indicating that there was sufficient reliability for comparative analysis. However, in a α -diversity analysis, there was no significant difference in species richness and diversity between samples. The Firmicutes phylum was common in both regions. At the species level, Weissella kandleri dominated in central (46.5%) and southern (30.8%) regions. Linear discriminant analysis effect size (LEfSe) analysis was performed to identify biomarkers representing the microbial community in each region. The LEfSe results pointed to statistically significant differences between the two regions in community composition, with Leuconostocaceae (71.4%) dominating in the central region and Lactobacillaceae (61.0%) dominating in the southern region. Based on these results, it can be concluded that the microbial communities of kimchi are significantly influenced by regional properties and that it can provide more useful scientific data to study the relationship between regional characteristics of kimchi and their microbial distribution.

• Proceedings of KOSOMES biannual meeting
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• 2017.11a
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• pp.209-209
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• 2017

• 축산식품과학과 산업
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• v.9 no.2
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• pp.26-35
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• 2020

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.312-325
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• 2015

In this review, we summarized the trends of genomics and transcriptomics research on peach, a model species of Rosaceae. Peach genome maps have been developed from various progeny groups with many next-generation sequencing (NGS) based single nucleotide polymorphism markers. Molecular markers of qualitative traits and quantitative trait loci (QTL) such as fruit characteristics, blooming date, and disease resistance have been analyzed. Among many characteristics, markers related to flesh softening and flesh adhesion are useful for marker assisted selection. Through comparative genomics, peach genome has been compared to the genome of Arabidopsis, Populus, Malus, and Fragaria species. Through transcriptomics and proteomics, fruit growth and development, and flavonoid synthesis, postharvest related transcriptomes and disease resistance related proteins have been reported. Recently, development of NGS based markers, construction of core collection of germplasm, and genotyping of various progenies have been preceded. In the near future, accurate QTL analysis and identification of useful genes are expected to establish a foundation for effective molecular breeding.

• Korean Journal of Veterinary Service
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• v.39 no.1
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• pp.41-49
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• 2016

In the present study, fast and robust methods for the next generation sequencing (NGS) were developed for analysis of PRRSV full genome sequences, which is a positive sensed RNA virus with a high degree of genetic variability among isolates. Two strains of PRRSVs (VR2332 and VR2332-R) which have been maintained in our laboratory were used to validate our methods and to compare with the sequence registered in GenBank (GenBank accession no. EF536003). The results suggested that both of strains had 100% coverage with the reference; the VR2332 had the coverage depth from minimum 3 to maximum 23,012, for the VR2332-R from minimum 3 to maximum 41,348, and 22,712 as an average depth. Genomic data produced from the massive sequencing capacities of the NGS have enabled the study of PRRSV at an unprecedented rate and details. Unlike conventional sequence methods which require the knowledge of conserved regions, the NGS allows de novo assembly of the full viral genomes. Therefore, our results suggested that these methods using the NGS massively facilitate the generation of more full genome PRRSV sequences locally as well as nationally in regard of saving time and cost.

• Korean Journal of Breeding Science
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• v.50 no.4
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• pp.424-433
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• 2018

Watermelon (Citrullus lanatus) is an economically important vegetable crop all over the world, which has functional compounds such as lycopene and citrulline. Gummy stem blight caused by Didymella bryoniae is one of the most devastative diseases in watermelon. Single nucleotide polymorphisms (SNPs), which are genetic variations occurring between individuals with respect to a single base, were often used to construct genetic linkage maps and develop molecular markers linked to a variety of horticultural traits and resistance to several diseases. In this study, we developed high-resolution melting (HRM) markers based on SNPs generated from NGS resequencing of two parents in watermelon. Plant materials were C. lanatus '920533' (female and susceptible parent), C. amarus 'PI 189225' (male and resistant parent), and their $F_1$ and $F_2$ progenies. A total of 13.6 Gbp ('920533') and 13.1 Gbp ('PI 189225') of genomic sequences were obtained using NGS analysis. A total of 6.09 million SNPs between '920533' and 'PI 189225' were detected, and 354,860 SNPs were identified as potential HRM primer sets. From these, a total of 330 primer sets for HRM analysis were designed. As a result, a total of 61 HRM markers that have polymorphic melting curves were developed. These HRM markers can be used for the construction of SNP-based linkage maps and for the analysis of quantitative trait loci (QTLs) related to gummy stem blight resistance.

• Journal of Korean Society of Forest Science
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• v.106 no.2
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• pp.121-129
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• 2017

The soil microbiome plays important roles in material cycling and plant growth in forest ecosystem. Although a lot of researches on forest soil fungi in Korea have been performed, the studies on forest soil bacterial communities have been limited. In this study, we conducted next generation sequencing (NGS) targeting 16S rRNA gene to investigate the soil bacterial communities from natural red pine (Pinus densiflora) forest in Mt. Janggunbong, Bonghwa-gun, Gyeongbuk, Korea. Our results showed that the entire bacterial communities in the study sites include the phyla Proteobacteria, Acidobacteria, Actinobacteria, Planctomycetes, which have been typically observed in forest soils. The composition ratio of Proteobacteria was the highest in the soil bacteria community. The results reflect that Proteobacteria is copiotroph, which generally favors relatively nutrient-rich conditions with abundant organic matter. Some rhizobia species such as Burkholderia, Bradyrhizobium, Rhizobium, which are known to contribute to soil nitrogen-fixation, exist in the study sites. As a result of correlation analysis between soil physicochemical characteristics and bacteria communities, the soil pH was significantly correlated with the soil bacteria compositions.