• Title/Summary/Keyword: Primer combinations

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A STUDY ON THE ADHESIVE BOND STRENGTH OF COMPOSITE RESIN TO Au-Ag-Cu-Pd ALLOY (Au-Ag-Cu-Pd합금과 복합레진간의 접착결합강도에 관한 연구)

  • Seol Young-Hoon;Jung Chang-Mo;Jeon Young-Chan
    • The Journal of Korean Academy of Prosthodontics
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    • v.32 no.3
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    • pp.378-395
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    • 1994
  • The purpose of this study was to investigate the effect of various metal surface treatments and adhesive systems on the flexural bond strength of composite resin to Au-Ag-Cu-Pd alloy. The specimens were divided into nine groups by the combinations of surface treatment methods and adhesive systems. The types of surface treatment in this study were alumina blasting only, alumina blasting-Sn plating, alumina blasting-heating and three kinds of adhesive system used in this study were Silicoater system(Heraeus Kulzer GmbH,Germany), Superbond C & B(Sun Medical Co.,Ltd.,Japan) and Cesead opaque primer(Kurary Co.,Ltd.,Japan). After surface treatments and adhesive systems were applied, each specimen was built up with Dentacolor composite resin (Heraeus Kulzer GmbH,Germany). Four-point flexural bond strength was measured by Instron universal testing machine (Model 4301,U.S.A.) and modes of failure were observed by SEM(JEOL,SSM-840A,Japan). The obtained results were as follows: 1. The group that was bonded with Superbond C & B after alumina blasting-heating shelved the highest bond strength with significant difference among the groups, except the group with Cesead opaque primer after alumina blasting-Sn plating(P<0.05). 2. In the groups bonded with Cesead opaque primer, there was significant difference only in the bond strength between the alumina blasting-Sn plating group and alumina blasting group, where the former showed a higher bond strength(P<0.05). 3. In the groups bonded with Silicoater system, there were no significant differences in bond strength regardless of the surface treatment method(P<0.05). 4. In SEM evaluation, the groups of high bond strength, especially bonded with Superbond C & B after alumina blasting-heating and Cesead opaque primer after alumina blasting-Sn plating, revealed mainly cohesive-adhesive failure, whereas the others showed the tendency of adhesive failure.

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cDNAs encoding the antigenic proteins in pathogenic strain of Entamoeba histolytica (이질아메바 병원성 분리주에서 발현되는 항원 단백질을 coding하는 cDNA)

  • 임경일;최종태
    • Parasites, Hosts and Diseases
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    • v.35 no.3
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    • pp.203-210
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    • 1997
  • The difrrrenlial display reverse transcription polymerase chain reaction (DDRT-PCR) aniilysis roils performed to identify the pathogellir strain specific amplicons. mRNAs were purified from the trophozoites of the pathogenif strain YS-27 and the non-pathogenic strain S 16. respectively. Three kinds of rirsl stranded rDNAs were reverse transcribed from the mRNAs by one base anchored oligo-dT 11M (M: A. C, or G) primers. Each cDNA lemplatr was used for DDRT-PCK analysis. A total of 144 pathogenic strain specific amplicons was observed in DDRT-PCR analysis using primer combinations of the 11 arbitrary primers and the 3 one base anchored oli해-dT11M primers. Of these 31 amplit'tons were verified as the amplirons amplified only from the mRNAs of the pathogenic strain by DNA slots biol llybridizatioil. Furthel cklaracleization of the 31 pathogenic strain sprcifil amplicons by DNA slot blot hybridlnation analysis using biotin labeled Probes or the PCR amplified DNA of rysteine proteinase genes revealed that 21 of them were amplliried from the maNAs of the cysteine proteinase genes. Four randomly selected amplirons out of the rest 10 amplirons were used fur screening of cDNA library followed by immunoscreening and all of them were turned outs to be amplified from the mRNA.

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Identification of Sex-specific Expression Markers in the Giant Tiger Shrimp (Penaeus monodon)

  • Khamnamtong, Bavornlak;Thumrungtanakit, Supaporn;Klinbunga, Sirawut;Aoki, Takashi;Hirono, Ikuo;Menasveta, Piamsak
    • BMB Reports
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    • v.39 no.1
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    • pp.37-45
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    • 2006
  • Bulked segregant analysis (BSA) and AFLP were used for isolation of genomic sex determination markers in Penaeus monodon. A total of 256 primer combinations were tested against 6-10 bulked genomic DNA of P. monodon. Five and one candidate female- and male-specific AFLP fragments were identified. Female-specific fragments were cloned and further characterized. SCAR markers derived from FE10M9520, FE10M10725.1, FE10M10725.2 and FE14M16340 provided the positive amplification product in both male and female P. monodon. Further analysis of these markers using SSCP and genome walk analysis indicated that they were not sex-linked. In addition, sex-specific (or differential) expression markers in ovaries and testes of P. monodon were analyzed by RAP-PCR (150 primer combinations). Twenty-one and fourteen RAP-PCR fragments specifically/differentially expressed in ovaries and testes of P. monodon were successfully cloned and sequenced. Expression patterns of 25 transcripts were tested against the first stranded cDNA of ovaries and testes of 3-month-old and broodstock-sized P. monodon (N = 5 and N = 7 - 10 for females and N = 4 and N = 5 - 7 for males, respectively). Five (FI-4, FI-44, FIII-4, FIII-39 and FIII-58) and two (M457-A01 and MII-51) derived RAP-PCR markers revealed female- and male-specific expression patterns in P. monodon. Surprisingly, MII-5 originally found in testes showed a higher expression level in ovaries than did testes of juvenile shrimps but a temporal female-specific pattern in P. monodon adults.

Ordered Differential Display from Cryphonectria parasitica

  • Kang, Hyun-Seok;Choi, Jin-Won;Park, Seung-Moon;Cha, Byeong-Jin;Yang, Moon-Sik;Kim, Dae-Hyuk
    • The Plant Pathology Journal
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    • v.16 no.3
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    • pp.142-146
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    • 2000
  • Ordered differential display using RT-PCR (ODD-PCR) was conducted to have a profile of the differently expressed genes between a hypovirulent strain of Cryphonectria parasitica (UEP1) and its isogenic wild type strain (EP155/2). ODD-PCR has advantages of high sensitivity, reproducibility, proportional representation, and limited number of primer combinations comparing with other differential display methods. RNAs were prepared from 1 and 5 day liquid culture of both hypovirulent and wild type strains, and were further evaluated with the marker genes of C. parasitica such as cryparin and mating factor MF2-1, which were already proven to be specifically down-regulated by the presence of mycovirus CHV1-713. ODD-PCR was conducted using those RNAs and expressed genes were categorized to five groups according to their temporal and quantitative expression patterns. Those fives groups are CPC, CPE, CPL, CPD, and CPU which represent constitutively-expressed, early-expressed, late-expressed, down-regulated, and up-regulated, respectively. Ninety two primer combinations out of a total of 192 have been tested so far. Among the twenty to fifty distinct bands per each reaction, an average of four to ten genes was identified as viral-regulated fungal genes. Those viral-specifc genes were further analyzed by DNA sequencing followed by homology search. Characterization of 30 clones including all five groups were conducted as a preliminary data and more are under investigation.

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Reverse Random Amplified Microsatellite Polymorphism Reveals Enhanced Polymorphisms in the 3' End of Simple Sequence Repeats in the Pepper Genome

  • Min, Woong-Ki;Han, Jung-Heon;Kang, Won-Hee;Lee, Heung-Ryul;Kim, Byung-Dong
    • Molecules and Cells
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    • v.26 no.3
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    • pp.250-257
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    • 2008
  • Microsatellites or simple sequence repeats (SSR) are widely distributed in eukaryotic genomes and are informative genetic markers. Despite many advantages of SSR markers such as a high degree of allelic polymorphisms, co-dominant inheritance, multi-allelism, and genome-wide coverage in various plant species, they also have shortcomings such as low polymorphic rates between genetically close lines, especially in Capsicum annuum. We developed an alternative technique to SSR by normalizing and alternating anchored primers in random amplified microsatellite polymorphisms (RAMP). This technique, designated reverse random amplified microsatellite polymorphism (rRAMP), allows the detection of nucleotide variation in the 3' region flanking an SSR using normalized anchored and random primer combinations. The reproducibility and frequency of polymorphic loci in rRAMP was vigorously enhanced by translocation of the 5' anchor of repeat sequences to the 3' end position and selective use of moderate arbitrary primers. In our study, the PCR banding pattern of rRAMP was highly dependent on the frequency of repeat motifs and primer combinations with random primers. Linkage analysis showed that rRAMP markers were well scattered on an intra-specific pepper map. Based on these results, we suggest that this technique is useful for studying genetic diversity, molecular fingerprinting, and rapidly constructing molecular maps for diverse plant species.

COMPARATIVE STUDY OF SHEAR BOND STRENGTH BETWEEN CP-TI/CO-CR ALLOY AND COMPOSITE RESINS

  • Yoon, Se-Hee;Pae, Ahran;Lee, Seok-Hyung;Lee, Ho-Rim
    • The Journal of Korean Academy of Prosthodontics
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    • v.45 no.6
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    • pp.805-814
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    • 2007
  • Statement of problem. Composite resin-veneered metal restorations can be used as an alternative to porcelain-fused-metal restorations. But, because of the relatively low bond strength of veneering composite to metal framework, various surface treatment methods have been introduced to improve the bond strength. Purpose. The object of this study was to compare the shear bond strength of different combinations of each of the two bonding systems and each of the two composite veneering resins to cp-Ti/Co-Cr alloy. Material and methods. Two resin bonding systems (metal conditioner containing MEPS monomer, tribochemical silicoating system) and two composite resins (Gradia, Sinfony) were tested on cp-Ti and Co-Cr alloy. Then, according to manufacturers' instructions, resin bonding systems and composite resins were applied. All test specimens were divided into four groups for each alloy; I) sandblast + Metal Primer II + Gradia (MG), II) sandblast + Metal Primer II + Sinfony (MS), III) Rocatec + Gradia (RG), IV) Rocatec + Sinfony (RS). The shear bond strength was determined using a universal testing machine and all data were statistically analyzed with Mann-Whitney test and Kruskal-Wallis test at the significance level of 0.05. Results. The mean (standard deviations) of shear bond strength according to the combinations of two bonding systems and two composite resins to cp-Ti arranged from 16.44 MPa to 17.07 MPa and the shear bond strength to Co-Cr alloy ranged from 16.26 MPa to 17.70 MPa. The result shows that the difference were not statistically significant. Conclusion. The shear bond strengths of composite resins to both cast cp-Ti and Co-Cr alloy were not significantly different between the metal conditioner and the tribochemical silicoating system. And no differences in bond strength were found between cp-Ti and Co-Cr alloy.

Selection and Genetic Relationship of Salt Tolerant Rice Mutants by in vitro Mutagenesis

  • Song, Jae Young;Kim, Dong Sub;Lee, Myung-Chul;Lee, Kyung Jun;Kim, Jin-Baek;Kim, Sang Hoon;Yun, Song Joong;Kang, Si-Yong
    • Journal of Radiation Industry
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    • v.4 no.4
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    • pp.307-312
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    • 2010
  • Plants have evolved physiological, biochemical and metabolic mechanisms to increase their survival under the adverse conditions. This present study has been performed to select salt-tolerant rice mutant lines through in vivo and in vitro mutagenesis with gamma-rays. For the selection of the salt-tolerant rice mutants, we conducted three times of selection procedure using 1,500 gamma ray mutant lines resulted from an embryo culture of the original rice cv. Dongan (wild-type, WT): first, selection in the a nutrient solution with 171 mM NaCl; second, selection under in vitro condition with 171 mM NaCl; and third, selection in a reclaimed saline land. Based on a growth comparison of the entries, out of the mutant lines, two putative 2 salt tolerant (ST) rice mutant lines, ST-87 and ST-301, were finally selected. The survival rate of the WT, ST-87 and ST-301 were 36.6%, 60% and 66.3% after 7 days in 171 mM NaCl treatment, respectively. The WT and two salt tolerant mutant lines were used to analyze their genetic variations. A total of 21 EcoRI and Msel primer combinations were used to analyze the genetic relationship of among the two salt-tolerant lines and the WT using the ABI3130 capillary electrophoresis system. In the AFLP analysis, a total of 1469 bands were produced by the 21 primer combinations, and 700 (47.6%) of them were identified as having polymorphism. The genetic similarity coefficients were ranged from 0.52 between the ST-87 and WT to 0.24 between the ST-301 and the WT. These rice mutant lines will be used as a control plot for physiological analysis and genetic research on salt tolerance.

A TILLING Rice Population Induced by Gamma-ray Irradiation and its Genetic Diversity

  • Cho, Hyun Yong;Park, Seo Jung;Kim, Dong Sub;Jang, Cheol Seong
    • Korean Journal of Breeding Science
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    • v.42 no.4
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    • pp.365-373
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    • 2010
  • TILLING (Targeting Induced Local Lesions IN Genomes) is broadly regarded as an excellent methodology for reverse genetics applications. Approximately 15,000 $M_3$ TILLING lines have been developed via the application of gamma-ray irradiation to rice seeds (cv. Donganbyeo), followed by subsequent selections. In an effort to evaluate the genetic diversity of the TILLING population, we have employed the AFLP multiple dominant marker technique. A total of 96 (0.64%) TILLING lines as well as Donganbyeo were selected randomly and their genetic diversity was assessed based on AFLP marker polymorphisms using 5 primer combinations. An average of 100.4 loci in a range of 97 to 106 was detected using these primer combinations, yielding a total of 158 (31.4%) polymorphic loci between Donganbyeo and each of the 96 lines. A broad range of similarity from 80% to 96% with an average of 89.4% between Donganbyeo and each of the 96 lines was also observed, reflecting the genetic diversity of the TILLING population. Approximately 28 polymorphic loci have been cloned and their sequences were BLAST-searched against rice whole genome sequences, resulting in 20 matches to each of the gene bodies including exon, intron, 1 kb upstream and 1 kb downstream regions. Six polymorphic loci evidenced changes in the coding regions of genes as compared to the rice pseudomolecules, 4 loci of which exhibited missense mutations and 2 loci of which exhibited silent mutations. Therefore, the results of our study show that the TILLING rice population should prove to be a useful genetic material pool for functional genomics as well as mutation breeding applications.

The effects of total-etch, wet-bonding, and light-curing of adhesive on the apical seal of a resin-based root canal filling system (접착제의 접착변수가 레진계 근관충전제의 근단밀폐효과에 미치는 영향)

  • Ryu, Won-Il;Shon, Won-Jun;Baek, Seung-Ho;Lee, In-Han;Cho, Byeong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.36 no.5
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    • pp.385-396
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    • 2011
  • Objectives: This study evaluated the effects of adhesion variables such as the priming concepts of canal wall and the curing modes of adhesives on the sealing ability of a resin-based root canal filling system. Materials and Methods: Apical microleakage of the Resilon-RealSeal systems filled with 3 different combinations of adhesion variables was compared with the conventional gutta-percha filling using a dye penetration method. Experimental groups were SEDC, Resilon (Resilon Research LLC) filling with self-etch RealSeal (SybronEndo) primer and dual-cure RealSeal sealer; NELC, Resilon filling with no etching, Scotchbond Multi-Purpose (3M ESPE) primer application and light-curing adhesive; and TELC, Resilon filling with Scotchbond Multi-Purpose primer and adhesive used under total etch / wet bonding and lightcure protocols. GPCS, gutta-percha filling with conventional AH26 plus sealer, was the control group. Results: The median longitudinal dye penetration length of TELC was significantly shorter than those of GPCS and SEDC (Kruskal-Wallis test, p < 0.05). In the cross-sectional microleakage scores, TELC showed significant differences from other groups at 2 to 5 mm from the apical foramen (Kruskal-Wallis test, p < 0.05). Conclusions: When a resin-based root canal filling material was used, compared to the self-etching primer and the dual-cure sealer, the total etch/wet-bonding with primer and light-curing of adhesive showed improved apical sealing and was highly recommended.

Simultaneous Detection of Four Foodborne Viruses in Food Samples Using a One-Step Multiplex Reverse Transcription PCR

  • Lee, Shin-Young;Kim, Mi-Ju;Kim, Hyun-Joong;Jeong, KwangCheol Casey;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.210-217
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    • 2018
  • A one-step multiplex reverse transcription PCR (RT-PCR) method comprising six primer sets (for the detection of norovirus GI and GII, hepatitis A virus, rotavirus, and astrovirus) was developed to simultaneously detect four kinds of pathogenic viruses. The size of the PCR products for norovirus GI and GII, hepatitis A virus (VP3/VP1 and P2A regions), rotavirus, and astrovirus were 330, 164, 244, 198, 629, and 449 bp, respectively. The RT-PCR with the six primer sets showed specificity for the pathogenic viruses. The detection limit of the developed multiplex RT-PCR, as evaluated using serially diluted viral RNAs, was comparable to that of one-step single RT-PCR. Moreover, this multiplex RT-PCR was evaluated using food samples such as water, oysters, lettuce, and vegetable product. These food samples were artificially spiked with the four kinds of viruses in diverse combinations, and the spiked viruses in all food samples were detected successfully.