• Title/Summary/Keyword: Bead-on-plate

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Modified CTAB DNA Methods for efficient DNA extraction from Rice (Oryza sativa L.) (벼 분자육종을 위한 CTAB DNA 추출 시스템 개량)

  • Lee, Jong-Hee;Kwak, Do-Yeon;Yeo, Un-Sang;Kim, Choon-Song;Jeon, Myeong-Gi;Kang, Jong-Rae;Park, Dong-Soo;Shin, Mun-Sik;Oh, Byeong-Geun;Hwang, Hung-goo
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.286-290
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    • 2008
  • Many important traits have been tagged allowing plant breeders to apply marker assisted selection (MAS) in rice. PCR itself is simple to set up, and requires little hands-on time. However, a crucial limiting step of MAS programs is the reliable and efficient extraction of DNA which can be performed on thousands of individuals. In this study, We describe a modification of the DNA extraction method, in which cetyltrimethylammonium bromide (CTAB) is used to extract DNA from leaf tissues for suitable MAS in rice. We followed the standard 2% CTAB extraction method in all the procedure. In addition we used the 1.2 ml 8-strip tube instead of 1.5 ml E-tubes to fit the 8-multichannel pipette and employ the 96 well plate to use the swing bucket centrifuge. Our modified CTAB DNA extraction method offers several advantages with respect to traditional and simple methods. 1) adult leaf samples collected in paddy field are applicable. 2) 96 leaf samples can be homogenized only one-time by using tungsten carbonate bead and 96well block. 3) semiautomatic loading method using 8-multichannel pipette from DNA extraction to electrophoresis of PCR products. 4) our system can extract about 400 leaf samples per day by only one technicion. Therefore, this method could be useful for marker assisted breeding in rice.

Intraerythrocytic culture and development of serological diagnostic tests of Babesia gibsoni 1. Indirect fluorescent antibody test and enzyme-linked immunosorbent assay for antibody detection of Babesia gibsoni infections in dogs (Babesia gibsoni의 적혈구내 배양법과 진단법 개발에 관한 연구 1. Babesia gibsoni 진단을 위한 간접형광항체법(IFAT)과 효소표지면역검사법(ELISA))

  • Suh, Myung-deuk;Shin, Yong-seung
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.583-593
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    • 1997
  • Indirect fluorescent antibody test(IFAT) and enzyme-linked imuunosorbent assay (IgG-ELISA) as serological diagnostic tools were conducted to evaluate the usefulness for diagnosis of canine babesiosis infected with Babesia gibsoni in domestic various dog breeds, american pit bullterrier, military shepherd, and mongrel dogs. The results obtained from this study were abstracted as follows. The nonionic detergent Triton X-100 and absorbent bio-bead $SM_2$ were useful reagents for the preparation of pure merozoite antigen of B gibsoni to be used in ELISA. The optimum reaction in ELISA was shown when the protein concentration of ELISA antigen was measured as 625ng/ml and the conjugate concentration was diluted into 1/6000 fold. The average OD value of ELISA in sera determined with negative responses in IFAT was measured as $0.255{\pm}0.051$(490nm) and the cut - off value of OD was determined as 0.399(490nm). The serum antibodies in both of IFAT and ELISA were detected on one week after artificially infected with B gibsoni and these high antibody titers, 512X in IFAT and 1024X in ELISA, were long lasted until 15 weeks after infection. The reproducibility of reaction and stability of the antigen absorbed microtitration polystyrene plate preserved in $4^{\circ}C$ refrigerator and $-20^{\circ}C$ freezer, respectively could be lasted until 135 days after storage. The positive rates in IFAT by dog breeds were shown 8.1%(60/744 heads) in mongrel dogs, 81.3%(78/96 heads) in american pit bullterrier and 15.6%(15/96 heads) in military shepherd, while the positive rate in ELISA shown 17.6%(131/744 heads) in mongrel dogs, 83.3%(80/96 heads) in american pit bullterrier and 36.5%(35/96 heads) in military shepherd, respiectively. In the total of 936 heads surveyed with IFAT and ELISA the positive rates in IFAT and ELISA were 16.4%(153/936 heads) and 26.3%(246/936 heads), respectivily. Agreement of reactions between IFAT and ELISA was shown 82.4% in 936 dog sera. The specificity and sensitivity of ELISA reaction were 83.5% and 76.5%, respectively. From the conclusion obtained in this study it was evaluated that IFAT and ELISA were useful as highly specific, sensitive and stable serelogical tools for the diagnosis of canine babesiosis in Korea.

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A Study on Material Characteristics and Manufacturing Techniques for Gold-granule Beads Excavated from the Neungsan-ri Temple Site in Buyeo (부여 능산리사지 출토 금제구슬의 재료학적 특성 및 제작기법 연구)

  • Yang, Soohyeon;Ro, Jihyun
    • Conservation Science in Museum
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    • v.26
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    • pp.67-82
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    • 2021
  • Two golden beads (Buyeo 5336) housed at the Buyeo National Museum were discovered in 1993 near the site of an ancient workshop in Neungsan-ri in Buyeo-gun, Chungcheongnam-do Province. These rare examples from the Baekje Kingdom of an application of granulation have maintained their original form intact, and thus serve as important materials for the investigation of production techniques applied. This study analyzed the composition of the golden beads using a portable X-ray fluorescence analyzer, a stereo microscope, and a scanning electron microscope with an energy dispersive X-ray spectrometer. The manufacturing technique was examined through the observation of the micro-shape and the surface condition and by a composition analysis of the joint part. In both beads, a hole was pierced in a hollow body and the bead was decorated with golden wires around the hole and gold granules in other parts. In some areas, golden granules had been attached to the gold plate and golden wires were then placed over the granules. The purity of both the wires and the granules was analyzed as 23.6 - 23.7K. A high copper content was detected in some of the parts where the granules were attached. The findings of a previous reproduction experiment and study of production methods suggest that the beads were made using the copper diffusion technique.