• Title/Summary/Keyword: Chain Reaction

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Sensitive method for the detection of Apple scar skin viroid(ASSVd) by nested reverse transcription-polymerase chain reaction

  • Lee, Sung-Joon;Kim, Chung;Sim, Sang-Mi;Lee, Dong-Hyuk;Lee, Jai-Youl
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.143.2-143
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    • 2003
  • A rapid and sensitive assay for the specific detection of plant viroids using reverse transcription-polymerase chain reaction(RT-PCR) has been developed already. The nested RT-PCR assay cloud be applied for the detection of apple scar skin viroid(ASSVd) from young leaves and other tissues. ASSVd has central conserved region(CCR), terminal left(T$\sub$L/) and terminal right(T$\sub$R/) domain. Primers were designed from these regions. Primer sets were successfully applicable for the amplification of full length or partial region of ASSVd by nested RT-PCR. Nested RT-PCR assay was more sensitive and accurate method to detect ASSVd from young trees during the early time of apple cultivation.

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Multiplex Reverse Transcription Polymerase Chain Reaction Assay for Simultaneous Detection of Five Cucurbit-infecting Viruses.

  • Lee, Su-Heon;Kim, Sang-Mok;Kim, Woo-Chang;Lee, Key-Woon
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.150.1-150
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    • 2003
  • A single-step multiplex reverse transcription polymerase chain reaction (RT-PCR) assay was developed for the simultaneous detection of five cucurbit-infecting viruses: cucumber mosaic virus (CMV), watermelon mosaic virus 2 (WMV2), zucchini yellow mosaic virus (ZYMV), cucumber green mottle mosaic virus (CGMMV), and kyuri green mottle mosaic virus (KGMMV). The multiplex RT-PCR provides a simple and rapid method for detecting various viruses in cucurbit plants, which will help diagnose many cucurbit plants at a time.

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Detection of Dirofilaria immitis by Reverse Transcription Polymerase Chain Reaction in Canine (역전사중합효소연쇄반응을 이용한 개심장사상충의 검출)

  • 이영준;박진호;권오덕;이주목
    • Journal of Veterinary Clinics
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    • v.16 no.1
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    • pp.177-181
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    • 1999
  • This study was undertaken to clarify the more accurate detecting method of Dirofilaria immitis. Seven dogs, average 7.47 years old, confirmed with Dirofilaria immitis infection by modified Knott's method were used as the experimental animals. cDNA was constructed using oligodT(15) primer after extracting total RNA from the blood of dogs that were confirmed with Dirofilaria immitis infection. As a result of polymerase chain reaction with template using constructed cDNA, the predicted products of a 378 base-pair DNA fragment was amplified. From these results, RT-PCR was more sensitive and effective than modified Knott's method to detect Dirofilaria immitis in dogs.

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Evaluation of Several Parameters of in situ Polymerase Chain Reaction (ISPCR) to Reduce the Leakage of Amplificants from Cells

  • Lee, Jae-Yung;Auh, Chung-Kyoon;George W. Jordan
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.70-76
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    • 2002
  • Proviral DNAs from HIV-1-infected CD4+ T cells (Molt/LAV cells) were amplified and detected in infected individual cells using polymerase chain reaction and in rifu hybridization. In this in situ PCR, three parameters were considered to achieve effective amplification and retention of amplificants inside the cells by making high molecular weight PCR products intracellularly, forming agarose matrix against the cells, and maintaining the appropriate PCR temperature profile. Over the cycles of ampliHcationl tailed primers with complementary overhanging sequences at their 5' sides manufactured high molecular weight products by using short primary products as a repeating unit. Agarose matrix could prevent the diffusion of the amplificants from the cells. Use of Thermanox coverslip inside the PCR tube offered target cells a similar temperature profile to that of conventional PCR in solution.

Identification and Differentiation of Cucumber Mosaic Virus Isolated from Forsythia koreana (CMV-Fk) Using PCR Techniques (PCR기법을 이용한 오이 모자이크 바이러스 개나리 분리주(CMV-Fk)의 동정과 구분)

  • 이상용;박선정;최장경
    • Korean Journal Plant Pathology
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    • v.14 no.4
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    • pp.308-313
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    • 1998
  • Reverse transcription and polymerase chain reaction (RT-PCR) techiniques were used to identification and differentiation of cucumber mosaic virus isolated from Forsythia koreana (CMV-Fk). RT-PCT used by two set of 20-mer primers one was CMV-common primers and another was CMV subgroup I-specific primers designed in a conserved region of the 3' end of CMV RNA3, amplified about 490 bp and 200 bp DNA fragments from CMV-Fk, respectively. CMV could be detected by RT-PCR at a dilution as low as 10-4 in forsythia crude sap extracts. Restriction enzyme analysis of RT-PCR products using EcoRI and MspI showed that CMV-Fk belonged to CMV subgroup I. But, analysis of RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR) showed heterogeneity of RNA3 between CMV-Fk and CMV-Y as a member of subgroup I.

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Random Isolation of Cadmium-induced Gene by Reverse Transcriptase PCR in Schizosaccharomyces pombe

  • Choi, Soon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.4 no.1
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    • pp.20-23
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    • 1994
  • The polymerase chain reaction was used to study the genes inducible under stress from the heavy metal cadmium. Schizosaccharomyces pombe, grown in the presence or absence of sublethal concentration of cadmium, was isolated to purify the total RNAs. The Induced RNA Random Fishing (IRRF) method in which random oligonucleotides were used as primers was applied to the identification of cadmium-induced gene expressions. A PCR-DNA product of 400-bp was cloned and sequenced. Computer analysis showed that this DNA has no homology with any known DNA sequences in GenBank or EMBL databases. The induction of this gene was confirmed by Northern blot analysis of total RNAs isolated from both cadmium-treated and untreated yeast cells.

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Cell- and Stage-Specific Expression of the Murine nm23-M5 Gene during Late Spermatogenesis and Spermiogenesis

  • Hwang Gyu-Chan;Ok Do-Won;Lee Mi-Suk;Kim Jin-Hoe
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.5-5
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    • 2002
  • Nucleoside diphosphate kinases (NDPKs) are conserved through evolution and have been shown to be involved in various biological phenomena. By functional screening in yeast, we identified a new member of the NDPK family, ㎚23-M5, which encodes a 211-amino acid protein with 86% identify to the human homolog, ㎚23-H5. Northern blot analysis reveals that ㎚23-M5 encodes two transcripts of 0.8 and 0.7 kb, which are highly and specifically expressed in adult testis. Reverse transcriptase polymerase chain reaction analysis shows that nm23-M5 first appears in pachytene spermatocytes and increase chain reaction in abundance through subsequent stages. (omitted)

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PCR-based Identification of Aflatoxigenic Fungi Associated with Iranian Saffron

  • Noorbakhsh, Reihaneh;Bahrami, Ahmad Reza;Mortazavi, Seyed Ali;Forghani, Bita;Bahreini, Maesoomeh
    • Food Science and Biotechnology
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    • v.18 no.4
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    • pp.1038-1041
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    • 2009
  • Aflatoxins are secondary metabolites produced by the aflatoxigenic fungi in suitable conditions. Saffron, Crocus sativus, is the most expensive spice in the world. Saffron is normally contaminated with soil and hand microflora during harvest and post-harvest operations. In this study, rapid assessment of aflatoxigenic fungi in saffron was accomplished using polymerase chain reaction. In total, 37 market samples were assayed in order to isolate aflatoxin-producing fungi. The 18.9% of the total samples were contaminated with aflatoxigenic fungi. Our results also show that most of the isolated fungi were saprophytes which are normally originated from soil during harvest and postharvest process.

Analysis of the DNA Fingerprints from the Teeth -Using Chelex$^\textregistered$ 100 as a Medium of Simple Extraction of DNA from the Teeth- (치아에서의 DNA 유전자지문 분석 -Chelex$^\textregistered$ 100을 매개체로 한 DNA추출-)

  • Chang-Lyuk Yoon
    • Journal of Oral Medicine and Pain
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    • v.20 no.2
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    • pp.515-528
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    • 1995
  • The human genomic deoxyribonucleic acid(DNA) was extracted from the pulp, dentin of 22 teeth by clelex, phenol methods. Samples of the tooth-derived DNA were amplified by polymerase chain reaction(PCR), electrophosed for sex determination by detection of X-Y homologus amelogenin gene and D1S80 locus detection The following results have been achieved. 1. Chelex and phenol method are effective to sex determination in the pulp and dentin 2. Chelex method is not suitable for detection of D1S80 locus. 3. Concentration and purity of DNA for teeth using chelex method is lower than using phenol method. From the above investigation, chelex method is simple, rapid for sex determination, but it is not suitable for detection of VNTRs.

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Comparison of Effects of Ultraviolet and $^{60}$ Co Gamma Ray Irradiation on Nylon 6 Mono-filaments

  • Ohtsuka, Mika;Suzuki, Yoshino;Sakai, Tetsuya;Netravali, Anil N.
    • Fibers and Polymers
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    • v.5 no.3
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    • pp.225-229
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    • 2004
  • The effect of UV and $^60{Co}$ gamma radiations on the physical and mechanical properties of nylon 6 mono-filaments with different draw ratios has been studied. Specimens were exposed to either up to 25 Mrad of gamma or up to 168 hrs of intense UV irradiation. The results show that nylon mono-filaments exposed to gamma rays, with much higher quantum energy than UV, undergo a larger extent of molecular chain scission. Higher irradiation dose also results in the production of insoluble, macroscopic three-dimensional cross-linked network structure. The amorphous regions with a lower density of cohesive energy (lower molecular orientation) show a higher extent of cross linking reaction whereas amorphous regions with a higher density of cohesive energy (higher orientation) show higher extent of chain scission reaction, irrespective of UV ray or gamma ray irradiation.