• Title/Summary/Keyword: SSU

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Optimization of DNA Extraction from a Single Living Ciliate for Stable and Repetitive PCR Amplification

  • Kim, Se-Joo;Min, Gi-Sik
    • Animal cells and systems
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    • v.13 no.3
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    • pp.351-356
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    • 2009
  • Ciliates are undoubtedly one of the most diverse protozoans that play a significant role in ecology. However, molecular examination, based on comparing the DNA sequences, has been done on a limited number of the species. Because most ciliates are uncultivable and their population sizes are often too small, it is usually difficult to obtain sufficient genomic DNA required for PCR based experiments. In the present study, we evaluated the effectiveness of four commercial DNA extraction procedures that extract high quality genomic DNA from a single ciliate cell. It was discovered that RED Extract-N-$Amp^{TM}$ PCR kit is the best method for removing PCR-inhibiting substances and minimizing DNA loss during purification. This method can also amplify more than 25 reactions of PCR. In addition, this technique was applied to single cells of 19 species belonged to 7 orders under 5 classes that isolated from mixed natural populations. Their small subunit ribosomal DNA (SSU rDNA) was successfully amplified. In summary, we developed a simple technique for the high-yield extraction of purified DNA from a single ciliate cell that may be more useful for rare ciliates, such as tiny and uncultivable marine microbes.

Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum

  • Yu, Jae-Ran;Lee, Soo-Ung;Park, Woo-Yoon
    • Parasites, Hosts and Diseases
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    • v.47 no.3
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    • pp.293-297
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    • 2009
  • Improved methods for detection of Cryptosporidium oocysts in environmental and clinical samples are urgently needed to improve detection of cryptosporidiosis. We compared the sensitivity of 7 PCR primer sets for detection of Cryptosporidium parvum. Each target gene was amplified by PCR or nested PCR with serially diluted DNA extracted from purified C. parvum oocysts. The target genes included Cryptosporidium oocyst wall protein (COWP), small subunit ribosomal RNA (SSU rRNA), and random amplified polymorphic DNA. The detection limit of the PCR method ranged from $10^3$ to $10^4$ oocysts, and the nested PCR method was able to detect $10^0$ to $10^2$ oocysts. A second-round amplification of target genes showed that the nested primer set specific for the COWP gene proved to be the most sensitive one compared to the other primer sets tested in this study and would therefore be useful for the detection of C. parvum.

Creep-Life Prediction and Standard Error Analysis of Type 316LN Stainless Steel (Type 316LN 스테인리스 강의 크리프 수명 예측과 표준오차 분석)

  • Yun S.N.;Kim W.G.;Liu W.S.;Yi W.
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2005.06a
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    • pp.1406-1411
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    • 2005
  • The creep rupture data for type 316LN stainless steels were collected through literature survey or experimental data produced in KAERI. Using these data, polynomial equations for predicting creep life were obtained by Larson-Miller (L-M), Orr-Sherby-Dorn (O-S-D) and Manson-Haferd (M-H) etc. time-temperature parametric (TTP) methods. Standard error of estimate (SEE) values for the each parameter was obtained with different temperatures through the statistical process of the creep data. The results of L-M, O-S-D and M-H methods showed good creep-life prediction, but M-H method showed better agreement than L-M and O-S-D methods. Especially, it was found that SEE values of M-H method at $700^{\circ}C$ were lower than that of L-M and O-S-D methods.

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First Record of Two Euplotes Ciliates (Ciliophora: Spirotrichea: Euplotida) from Korea

  • Park, Mi-Hyun;Kim, Se-Joo;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.26 no.1
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    • pp.21-27
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    • 2010
  • Two marine euplotid ciliates, i.e. Euplotes cristatus Kahl, 1932 and E. minuta Yocom, 1930, were collected from the public waterfront of Incheon on the Yellow Sea and from the Songjeong Beach, Busan, in the Strait of Korea, respectively. These two species were verified as unrecorded species in Korea. These species were described based on live observation, protargol impregnation, and silver nitrate impregnation. In addition, the small subunit ribosomal DNA (SSU rDNA) sequences of the two species were compared with previously known sequences of the Euplotes species. Euplotes cristatus has an elongated oval form, size in vivo of $60-84{\times}38-68\;{\mu}m$, 35-50 adoral zone of membranelles (AZM), 10 frontoventral cirri (FVC), 5 transverse cirri (TC), 4-5 caudal cirri (CC), 8 dorsal kineties (DK), 10-16 dorsal cilia of middle DK, and silverline system of single-vannus type. Euplotes minuta has a small ovoid form ($44-53{\times}26-35\;{\mu}m$ in vivo), 31-41 AZM, 10 FVC, 5 TC, 4 CC, 9 DK, 10-12 dorsal cilia of middle DK, and silverline system of single-vannus type.