• Title/Summary/Keyword: Molecular identification

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Molecular Identification of Reynoutria japonica Houtt. and R. sachalinensis (F. Schmidt) Nakai Using SNP Sites

  • Park, Hana;Yoon, Chang Young;Kim, Jin Sook;Kim, Joo-Hwan
    • Korean Journal of Plant Resources
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    • v.28 no.6
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    • pp.743-751
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    • 2015
  • Reynoutria japonica and R. sachalinensis have been used as medicinal resources in Korea. However, it is difficult to identify and determine these medicinal herbs correctly because they are usually customized and purchased as the fragmented rhizomes types. To develop molecular markers for distinguishing two species, we analyzed and compared the chloroplast DNA sequences of seven loci (atpB, matK, ccD-psaI, atpF-H, trnL-trnF, psbK-I and rpl32-trnL). Among them, we found two effective SNPs in psbK-I region for R. japonica and atpF-H region for R. sachalinensis. Based on these SNP sites, we designed the new R. japonica- specific primer which is able to amplify 300 bp fragment in psbK-I region. A similar strategy was applied for the atpF-H region of R. sachalinensis. These molecular markers would be successfully applied to recognize R. japonica and R. sachalinensis.

Comparison of digital PCR platforms using the molecular marker

  • Cherl-Joon Lee;Wonseok Shin;Minsik Song;Seung-Shick Shin;Yujun Park;Kornsorn Srikulnath;Dong Hee Kim;Kyudong Han
    • Genomics & Informatics
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    • v.21 no.2
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    • pp.24.1-24.7
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    • 2023
  • Assays of clinical diagnosis and species identification using molecular markers are performed according to a quantitative method in consideration of sensitivity, cost, speed, convenience, and specificity. However, typical polymerase chain reaction (PCR) assay is difficult to quantify and have various limitations. In addition, to perform quantitative analysis with the quantitative real-time PCR (qRT-PCR) equipment, a standard curve or normalization using reference genes is essential. Within the last a decade, previous studies have reported that the digital PCR (dPCR) assay, a third-generation PCR, can be applied in various fields by overcoming the shortcomings of typical PCR and qRT-PCR assays. We selected Stilla Naica System (Stilla Technologies), Droplet Digital PCR Technology (Bio-Rad), and Lab on an Array Digital Real-Time PCR analyzer system (OPTOLANE) for comparative analysis among the various droplet digital PCR platforms currently in use commercially. Our previous study discovered a molecular marker that can distinguish Hanwoo species (Korean native cattle) using Hanwoo-specific genomic structural variation. Here, we report the pros and cons of the operation of each dPCR platform from various perspectives using this species identification marker. In conclusion, we hope that this study will help researchers to select suitable dPCR platforms according to their purpose and resources.

Identifications of Predominant Bacterial Isolates from the Fermenting Kimchi Using ITS-PCR and Partial 16S rDNA Sequence Analyses

  • CHIN HWA SUP;BREIDT FRED;FLEMING H. P.;SHIN WON-CHEOL;YOON SUNG-SIK
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.68-76
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    • 2006
  • Despites many attempts to explore the microbial diversity in kimchi fermentation, the predominant flora remains controversial to date. In the present study, major lactic acid bacteria (LAB) were investigated in Chinese cabbage kimchi in the early phase of fermention. For the samples over pH 4.0, viable cell counts of Leuconostoc and Pediococcus were $10^6\;cfu/ml$ and below $10^2\;cfu/ml$, respectively, and 20 isolates out of 172 were subjected to a biochemical identification (API 50 CH kit) as well as molecular-typing methods including ITSPCR with a RsaI digestion and 16s rRNA gene sequence analysis for species confirmation. Seven isolates were nicely assigned to Lb. brevis, 6 to Leuconostoc spp. (2 mesenteroides, 2 citreum, I carnosum, I gasicomitatum), 4 to Weissella (3 kimchii/cibaria, 1 hanii) and 2 to other Lactobacillus spp. (1 farciminis, 1 plantarum). On the other hand, the biochemical identification data revealed 9 strains of Lb. brevis, 6 strains of Leuconostocs,2 strains of Lb. plantarum and 1 strain each of Lb. coprophilus and Lactococcus lactis. However, a single isolates, YSM 16, was not matched to the ITS-PCR database constructed in the present study. Two Lb. brevis strains by API 50 CH kit were reassigned to W kimchii/cibaria, Lb. coprophilus or W hanii, respectively, judging from the results by the above molecular typing approaches. As a whole, the identification data obtained by the biochemical test were different from those of ITS-PCR molecular method by about $63\%$ at genus-level and $42\%$ at species-level. The data by the ITS-PCR method conclusively suggest that predominant LAB species is probably heterolactic Lb. brevis, followed by W kimchii/cibaria, Leuc. mesenteroides, and Leuc. citreum, in contrast to the previous reports [3] that Leuc. mesenteroides is the only a predominant species in the early phase kimchi fermentation.

Identification of Species and Sex of Korean Roe Deer (Capreolus pygargus tianschanicus) Using SRY and CYTB Genes

  • Han, Sang-Hyun;Cho, In-Cheol;Lee, Sung-Soo;Tandang, Leoncia;Lee, Hang;Oh, Hong-Shik;Kim, Byoung-Soo;Oh, Moon-You
    • Animal cells and systems
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    • v.11 no.2
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    • pp.165-168
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    • 2007
  • The nucleotide sequences of a male-specific marker sex determining region Y (SRY) gene and a mitochondrial cytochrome B (CYTB) gene were characterized and analyzed to establish a molecular method for identification of species and sex of Korean roe deer (Capreolus pygargus tianschanicus). Similarity search result of SRY sequences showed very similar result to those reported in Moose (Alces alces) and Reindeer (Rangifer tarandus), both of which had 95.9% similarity in identity. CYTB genes were very similar to those reported in Siberian roe deer (C. pygargus pygargus) which had 98.6% similarity and not to European roe deer (C. capreolus), suggesting that the DNA samples tested were of Siberian roe deer lineage. Polymerase chain reaction (PCR)-based sex typing successfully discriminated between carcasses of male and female roe deer. Males had SRY band on agarose gels and females did not. The result of this molecular sex typing provided similar information with that obtained by genital organ observation. Therefore, this molecular method using male specific marker SRY and mitochondrial CYTB genes would be very useful for identification of the species and sex of the carcass remains of roe deer.

Molecular and Morphological Identification of Ophichthid Leptocephali from the South Sea of Korea (남해에서 채집된 바다뱀과 엽상자어(Leptocephali)의 형태 및 분자 동정)

  • Ji, Hwan-Sung;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.22 no.4
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    • pp.279-284
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    • 2010
  • Four leptocephali (TL 109.8~129.7 mm) of the family Ophichthidae, collected from Sangju in the South Sea of Korea, were identified using morphological and molecular methods. Our four leptocephali were similar to Ophichthus sp. based on morphological characters: the melanophores present in series of eight gut swellings; total myomeres 144~151; body depth in total length less than 10%; eight or nine black horizontal bands present from anus to caudal fin base. On the other hand, our four leptocephali were identified to Pisodonophis sp. based on 826 base pairs of 12S rRNA gene nucleotide sequences. Our results showed that morphology-based identification did not agree with molecular identification, indicating difficulty in differentiating morphologically between Ophichthus and Pisodonophis leptocephali. We, herein, firstly describe morphological characteristics of Pisodonophis sp. leptocephali from Korea.

Validation of Morphology-based Identification of Two Cynoglossidae Larvae using Mitochondrial DNA (참서대과(Pisces: Cynoglossidae) 자어 2종의 미토콘드리아 DNA에 의한 형태동정의 타당성)

  • Kwun, Hyuck-Joon;Kim, Jin-Koo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.5
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    • pp.482-488
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    • 2010
  • Three specimens of Cynoglossidae larvae were collected from the southern Korean Sea in May and August of 2009, and were identified using morphological and molecular analysis. Specimens were divided into two groups based on the number of elongated dorsal fin rays on the top of the head: Cynoglossidae sp. A was defined as having two elongated dorsal fin rays, while Cynoglossidae sp. B possessed a single elongated dorsal fin ray. One specimen of Cynoglossidae sp. A, a post-larva with a notochord length (NL) of 5.8 mm was thought to be a Cynoglossus joyneri larva based on the presence of 115 dorsal pterogiophores, 85 anal pterogiophores, and 50 myomeres. Two specimens of Cynoglossidae sp. B, a 4.1 mm NL larva and a 11.3 mm NL juvenile, were thought to be Cynoglossus abbreviatus based on the presence of yolk in the former and 133 dorsal fin rays, 105 anal fin rays, and 63 myomeres in the latter. To test this morphology-based identification, molecular analysis was conducted using 419-422 bp of mitochondrial DNA 16S rRNA. Cynoglossidae sp. A was clearly matched to a Cynoglossus joyneri adult (d=0.000) and Cynoglossidae sp. B clustered closely with Cynoglossus abbreviatus adults (d=0.002). A neighbor-joining tree supported this robust relationship (bootstrap value=100%). Therefore, these molecular data validate the morphological identification of the two Cynoglossidae larval species.

What Determines Star Formation Rates?

  • Evans, Neal
    • The Bulletin of The Korean Astronomical Society
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    • v.41 no.2
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    • pp.29.4-29.4
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    • 2016
  • The relations between star formation and properties of molecular clouds are studied based on a sample of star forming regions in the Galactic Plane. Sources were selected by having radio recombination lines to provide identification of associated molecular clouds and dense clumps. Radio continuum and mid-infrared emission were used to determine star formation rates, while 13CO and submillimeter dust continuum emission were used to obtain masses of molecular and dense gas, respectively. We test whether total molecular gas or dense gas provides the best predictor of star formation rate. We also test two specific theoretical models, one relying on the molecular mass divided by the free-fall time, the other using the free-fall time divided by the crossing time. Neither is supported by the data. The data are also compared to those from nearby star forming regions and extragalactic data. The star formation "efficiency," defined as star formation rate divided by mass, spreads over a large range when the mass refers to molecular gas; the standard deviation of the log of the efficiency decreases by a factor of three when the mass of relatively dense molecular gas is used rather than the mass of all the molecular gas.

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