• 제목/요약/키워드: Molecular techniques

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Improved DNA Extraction Method for Molecular Diagnosis from Smaller numbers of Cells

  • Oh, Seo Young;Han, Jeong Yeon;Lee, So Ra;Lee, Hoon Taek
    • 대한임상검사과학회지
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    • 제46권3호
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    • pp.99-105
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    • 2014
  • Isolating total DNA from small samples using traditional methods is difficult and inefficient mainly due to loss of DNA during filtration and precipitation. With advances in molecular pathology, DNA extraction from micro-dissected cells has become essential in handling clinical samples. Genomic DNA extraction using small numbers of cells can be very important to successfully PCR amplify DNA from small biopsy specimens. We compared our experimental genomic DNA extraction method (A) with two other commercially available methods: using spin columns (B), and conventional resins (C), and determined the efficacy of DNA extraction from small numbers of cells smeared on a glass slide. Approximately 50, 100, 200, 500 and 1000 cells were isolated from fine needle aspiration biopsy (FNAB) slides aspirated from histologically proven papillary thyroid carcinoma masses. DNA was extracted using the three techniques. After measuring DNA quantity, PCR amplification was performed to detect the ${\beta}$-globin and $BRAF^{V600E}$ gene mutations. DNA extracted by method (A) showed better yield than the other methods in all cell groups. With our method, a suitable amount of genomic DNA to produce amplification was extracted from as few as 50 cells, while more than 100 to 200 cells were required when methods (B) or (C) were applied. Our genomic DNA extraction method provides high quality and improved yields for molecular analysis. It will be especially useful for paucicellular clinical samples which molecular pathologists often confront when handling fine needle aspiration cytology, exfoliative cytology and small biopsy specimens.

Analysis of silkworm molecular breeding potential using CRISPR/Cas9 systems for white egg 2 gene

  • Park, Jong Woo;Yu, Jeong Hee;Kim, Su-Bae;Kim, Seong-Wan;Kim, Seong-Ryul;Choi, Kwang-Ho;Kim, Jong Gil;Kim, Kee Young
    • International Journal of Industrial Entomology and Biomaterials
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    • 제39권1호
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    • pp.14-21
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    • 2019
  • Genome editing by CRISPR/Cas9, a third-generation gene scissor in molecular breeding at the genome level, is attracting much attention as one of the breeding techniques of the future. In this study, genetic and phenotypic analysis was used to examine the responsiveness of the Bakokjam variety of the silkworm Bombyx mori to molecular breeding using CRISPR/Cas9 in editing the white egg 2 (w-2) gene. The nucleotide sequence of the w-2 gene was analyzed and three different guide RNAs (gRNA) were prepared. The synthesized gRNA was combined with Cas9 protein and then analyzed by T7 endonuclease I after introduction into the Bm-N silkworm cell line. To edit the silkworm gene, W1N and W2P gRNA and Cas9 complexes were microinjected into silkworm embryos. Based on the results of microinjection, the hatching rate was 16-24% and the incidence of mutation was 33-37%. The gene mutation was verified in the heterozygous F1 generation, but no phenotypic change was observed. In F2 homozygotes generated by F1 self-crosses, a mutant phenotype was observed. These results suggest that silkworm molecular breeding using the CRISPR/Cas9 system is possible and will be a very effective way to shorten the time required than the traditional breeding process.

HgCdTe를 이용한 Infrared Detector의 제조와 특성 (Fabrication and Its Characteristics of HgCdTe Infrared Detector)

  • 김재묵;서상희;이희철;한석룡
    • 한국군사과학기술학회지
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    • 제1권1호
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    • pp.227-237
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    • 1998
  • HgCdTe Is the most versatile material for the developing infrared devices. Not like III-V compound semiconductors or silicon-based photo-detecting materials, HgCdTe has unique characteristics such as adjustable bandgap, very high electron mobility, and large difference between electron and hole mobilities. Many research groups have been interested in this material since early 70's, but mainly due to its thermodynamic difficulties for preparing materials, no single growth technique is appreciated as a standard growth technique in this research field. Solid state recrystallization(SSR), travelling heater method(THM), and Bridgman growth are major techniques used to grow bulk HgCdTe material. Materials with high quality and purity can be grown using these bulk growth techniques, however, due to the large separation between solidus and liquidus line on the phase diagram, it is very difficult to grow large materials with minimun defects. Various epitaxial growth techniques were adopted to get large area HgCdTe and among them liquid phase epitaxy(LPE), metal organic chemical vapor deposition(MOCVD), and molecular beam epitaxy(MBE) are most frequently used techniques. There are also various types of photo-detectors utilizing HgCdTe materials, and photovoltaic and photoconductive devices are most interested types of detectors up to these days. For the larger may detectors, photovoltaic devices have some advantages over power-requiring photoconductive devices. In this paper we reported the main results on the HgCdTe growing and characterization including LPE and MOCVD, device fabrication and its characteristics such as single element and linear array($8{\times}1$ PC, $128{\times}1$ PV and 4120{\times}1$ PC). Also we included the results of the dewar manufacturing, assembling, and optical and environmental test of the detectors.

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제주도 한라산에 서식하는 도입종 야생멧돼지에 대한 분자유전학적 분석 (A Molecular Genetic Analysis of the Introduced Wild Boar Species (Sus scrofa coreanus) on Mount Halla, Jeju Island, Korea)

  • 한상현;오장근;조인철;고문석;김태욱;장민호;김병수;박수곤;오홍식
    • 한국환경생태학회지
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    • 제25권5호
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    • pp.658-665
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    • 2011
  • 제주도에서는 절멸된 것으로 간주되었던 멧돼지들이 최근 한라산 인근지역에서 발견되었다. 본 연구는 분자유전학적 실험기법을 바탕으로 한라산 멧돼지들이 가축돼지들과 이종교배된 것들인지를 조사하였다. 또한 동일 종내에서의 유전적 유연관계와 분자 성판별을 시험하였다. 가축돼지 품종들(Landrace, Large White, Berkshire, Hampshire, Duroc)과의 교배여부는 핵 DNA와 미토콘드리아 DNA에서 4 종류의 분자 표지인자(MC1R, KIT, 조절영역, ND2)를 적용하여 시험하였다. 야생멧돼지 집단의 모든 개체들이 동일한 mtDNA 조절영역 서열을 나타내었고, 그 서열들은 중국 동북부 재래돼지들과 동일하였으나 기존에 보고된 한반도 멧돼지의 서열들과는 다른 것으로 확인되었다. 이상의 연구결과는 한라산 멧돼지집단이 중국 재래돼지 품종들과 근연이면서, 기존에 연구되지 않았던 유전적 계통에서 유래한 것으로 사료된다. 분자 성판별 결과 수컷에 비해 암컷이 2 배 이상으로 확인되어, 한라산 야생멧돼지 집단이 팽창하고 있으며, 조절하지 않으면 집단 규모는 극적으로 증가할 것이다.

마이크로기공 고분자(PIM-1)의 분자량 분포에 따른 이산화탄소 기체 분리막의 성능 변화 연구 (Effect of Molecular Weight Distribution of Intrinsically Microporous Polymer (PIM-1) Membrane on the CO2 Separation Performance)

  • 권지민;손혜정;김진욱;이창수
    • 멤브레인
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    • 제33권6호
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    • pp.362-368
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    • 2023
  • 이 연구 논문은 기후 변화에 대한 전 세계적인 우려와 온실 가스 배출 감소를 위한 필수적인 요구에 대응하여 마이크로기공 고분자(PIM-1)의 이용을 탐구한 것이다. 연구는 PIM-1 막을 이산화탄소(CO2) 가스 분리 막으로 사용하는 현대적인 소재로서의 응용에 집중하고 있다. 연구는 PIM-1 막의 합성, 분자량 제어, 그리고 제각각의 특성 분석 기술을 통해 포괄적인 통찰을 제공하며, 이러한 특성 분석 기술을 통해 PIM-1의 고유한 교차결합 및 강성 구조에서 비롯된 내재적 다공성이 특히 이산화탄소의 선택적 투과에 활용되고 있다. 논문은 PIM-1의 가교된 구조로부터 비롯된 내재적 다공성이 특히 이산화탄소의 선택적 투과에 활용되고 있다. 논문은 PIM-1의 튜닝 가능한 화학적 특성을 강조하며, 가스 분리 막의 맞춤 및 최적화를 가능케 하는 특성에 대한 이해를 제시하고 있다. 분자량을 통제함으로써 고분자량(H-PIM-1) 막은 낮은 분자량 대비 더 뛰어난 CO2 투과성과 선택성을 나타내며, 이를 통해 PIM-1 막의 특성을 조절하는 데 분자량의 중요성을 강조하고 있다. 연구 결과는 PIM-1 막 특성을 조절하는 데 분자량이 중요한 역할을 하는 것을 강조하며, 이는 기후 변화의 긴급한 글로벌 도전에 대응하기 위한 효율적이고 선택적인 CO2 포집을 위한 차세대 막 기술의 발전에 기여하고 있다.

3D 프린팅 소재 화학물질의 독성 예측을 위한 Data-centric XAI 기반 분자 구조 Data Imputation과 QSAR 모델 개발 (Data-centric XAI-driven Data Imputation of Molecular Structure and QSAR Model for Toxicity Prediction of 3D Printing Chemicals)

  • 정찬혁;김상윤;허성구;;신민혁;유창규
    • Korean Chemical Engineering Research
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    • 제61권4호
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    • pp.523-541
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    • 2023
  • 3D 프린터의 활용이 높아짐에 따라 발생하는 화학물질에 대한 노출 빈도가 증가하고 있다. 그러나 3D 프린팅 발생 화학물질의 독성 및 유해성에 대한 연구는 미비하며, 분자 구조 데이터의 결측치로 인해 in silico 기법을 사용한 독성예측 연구는 저조한 실정이다. 본 연구에서는 화학물질의 분자구조 정보를 나타내는 주요 분자표현자의 결측치를 보간하여 3D 프린팅의 독성 및 유해성을 예측한 Data-centric QSAR 모델을 개발하였다. 먼저 MissForest 알고리즘을 사용해 3D 프린팅으로 발생되는 유해물질의 분자표현자 결측치를 보완하였으며, 서로 다른 4가지 기계학습 모델(결정트리, 랜덤포레스트, XGBoost, SVM)을 기반으로 Data-centric QSAR 모델을 개발하여 생물 농축 계수(Log BCF)와 옥탄올-공기분배계수(Log Koa), 분배계수(Log P)를 예측하였다. 또한, 설명 가능한 인공지능(XAI) 방법론 중 TreeSHAP (SHapley Additive exPlanations) 기법을 활용하여 Data-centric QSAR 모델의 신뢰성을 입증하였다. MissForest 알고리즘 기반 결측지 보간 기법은, 기존 분자구조 데이터에 비하여 약 2.5배 많은 분자구조 데이터를 확보할 수 있었다. 이를 바탕으로 개발된 Data-centric QSAR 모델의 성능은 Log BCF, Log Koa와 Log P를 각각 73%, 76%, 92% 의 예측 성능으로 예측할 수 있었다. 마지막으로 Tree-SHAP 분석결과 개발된 Data-centric QSAR 모델은 각 독성치와 물리적으로 상관성이 높은 분자표현자를 통하여 선택함을 설명할 수 있었고 독성 정보에 대한 높은 예측 성능을 확보할 수 있었다. 본 연구에서 개발한 방법론은 다른 프린팅 소재나 화학공정, 그리고 반도체/디스플레이 공정에서 발생 가능한 오염물질의 독성 및 인체 위해성 평가에 활용될 수 있을 것으로 사료된다.

Molecular and Morphological Characterization of Green Mold, Trichoderma spp. isolated from Oyster Mushrooms

  • Choi, In-Young;Hong, Seung-Beom;Yadav, Mahesh C.
    • Mycobiology
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    • 제31권2호
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    • pp.74-80
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    • 2003
  • Isolates of Trichoderma spp. collected from Pleurotus ostreatus and P. eryngii beds, which included loosened substrate compactness and development of green colour, were grouped into three species. The occurrence of different species of Trichoderma was as T. cf. virens(70.8%), T. longibrachiatum(16.7%) and T. harzianum(12.5%). The conidia of Trichoderma spp. were ellipsoidal, obovoid and phialides were bowling pins, lageniform and the length of phialides was $3.5{\sim}10.0{\times}1.3{\sim}3.3{\mu}m$. Phialides of T. cf. virens and T. harzianum were tending clustered, but it was solitary disposition in T. longibrachiatum. T. cf. virens was characterized by predominantly effuse conidiation, sparingly branched, and fertile to the apex and it was penicillate type. RAPD analysis could detect variability amongst three different species of Trichoderma using two newly designed URP-primers. However, intra-specific variation could not be detected in all the isolates except for rDNA sequence data classified Trichoderma isolates into three distinct groups representing three species. The profiles of rDNA sequences of isolates representing a species showed high similarity in T. cf. virens and T. harzianum. However, there was a variation in rDNA sequences of isolates representing T. longibrachiatum. The results of present study reveals that molecular techniques of RAPD and rDNA sequencing can greatly aid in classification based on morphology and precise identification of fast evolving species of Trichoderma.

Synthesis and Characterization of a Tetrathiafulvalene-Based Polymer

  • Lee, Se-Hyun;Wang, Lei;Hwang, Seok-Ho;Lee, Myong-Hoon;Jeong, Kwang-Un
    • Bulletin of the Korean Chemical Society
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    • 제33권5호
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    • pp.1451-1456
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    • 2012
  • A novel tetrathiafulvalene (TTF)-based main-chain polymer (6TTF-polymer) was successfully synthesized via a condensation polymerization between a newly synthesized dihydroxy TTF derivative and a malonyl chloride, and its chemical structure was characterized by spectroscopic techniques. Molecular weight of the 6TTF-polymer (9,030 g/mol by gel permeation chromatography) was large enough to form the ductile film. The electrochemical and optical properties of the 6TTF-polymer were further estimated by cyclic voltammetry, ultraviolet and photoluminescence spectroscopes. The highest occupied molecular orbital level ($E_{HOMO}$=-4.79 eV) and band-gap energy ($E_g$=1.91 eV) of the 6TTF-polymer suggested that TTF-based polymer could act as a good electron donating material for the optoelectronic applications.

Synthesis, and Structural and Thermal Characterizations of Tetrasulfonated Poly(arylene biphenylsulfone ether) Copolymer Ion Conducting Electrolytes

  • Yoo, Dong-Jin;Hyun, Seung-Hak;Kim, Ae-Rhan;Kumar, G. Gnana;Nahm, Kee-Suk
    • Bulletin of the Korean Chemical Society
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    • 제32권11호
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    • pp.4041-4048
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    • 2011
  • High molecular weight tetrasulfonated poly(arylene biphenylsulfone ether) (TsPBPSEH) copolymers containing up to four pendant sulfonate groups per repeat unit were synthesized via aromatic nucleophilic displacement condensation from 4,4'-bis(4-chloro-3-sulfonatophenylsulfonyl)biphenyl-2,2'-disulfonate (SBCSBPD), 4,4'-dichlorodiphenylsulfone (DCDPS) and 4,4'-(hexafluoroisopropylidene)diphenol (6F-BPA). The synthesized copolymers were structurally characterized using $^1H$ NMR and FT-IR techniques. They were analytically pure, amorphous and were readily soluble in a wide range of organic solvents. Electrolyte membranes were successfully cast using the synthesized polymers with various sulfonation levels and N-methyl-2-pyrrolidinone. This new class of polymer membranes exhibited elevated thermal and physical stabilities and reduced swelling at high temperatures. An increase of acidic functional groups in the copolymer yielded high ion exchange capacity and moderate ionic conductivity values even at higher temperatures, which makes them potential ion conducting candidates.

Gd-Complexes of DTPA-bis(amides) Functionalized by Pyridine and Picolinamide: Synthesis, Thermodynamic Stability, and Relaxivity Properties

  • Sk, Nasiruzzaman;Park, Ji-Ae;Chang, Yong-Min;Kim, Tae-Jeong
    • Bulletin of the Korean Chemical Society
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    • 제29권6호
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    • pp.1211-1216
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    • 2008
  • A series of DTPA-bis(amides) functionalized by pyridine (1a-c) and N-phenylpicolinamide) (1d-e) and their Gd(III)-complexes of the type [Gd(1)($H_2O$)]·x$H_2O$ (2a-e) were prepared and characterized by analytical and spectroscopic techniques. Potentiality of 2a-e as contrast agents for magnetic resonance imaging (MRI CA) was investigated by measuring relevant physicochemical properties and relaxivities and compared with [Gd(DTPA-BMA)($H_2O$)] (DTPA-BMA=N,N''-di(methylcarbamoylmethyl)diethylenetriamine-N,N',N''-triacetate) ($Omniscan^{(R)}$). The R1 relaxivities of aqueous solutions of 2a-c are in the range of 3.33 -5.02 $mM^{-1}$$sec^{-1}$, which are comparable with those of $Omniscan^{(R)}$ (r1=4.58 $mM^{-1}sec^{-1}$). Complexes 2d-e, insoluble in water, exhibit relatively higher R1 values (8.1- 8.3 $mM^{-1}sec^{-1}$) in HP-$\beta$-CD solutions.