• Title/Summary/Keyword: universal primer

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Shear bond strength of Universal bonding systems to Ni-Cr alloy (니켈-크롬 합금에 대한 다용도 접착 시스템의 전단결합강도)

  • Song, So-Yeon;Son, Byung-Wha;Kim, Jong-Yeob;Shin, Sang-Wan;Lee, Jeong-Yol
    • The Journal of Korean Academy of Prosthodontics
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    • v.53 no.4
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    • pp.295-300
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    • 2015
  • Purpose: The aim of this study was to evaluate the shear bond strength between Ni-Cr alloy and composite resin using universal adhesive systems coMPared to conventional method using metal primers. Materials and methods: For this study, a total of 120 cast commercial Ni-Cr alloy (Vera Bond 2V) disks were embedded in acrylic resin, and their surfaces were smoothed with silicon carbide papers and airborne-particle abrasion. Specimens of each metal were divided into 6 groups based on the combination of metal primers (Metal primer II, Alloy primer, Metal & Zirconia primer, MKZ primer) and universal adhesive systems (Single Bond Universal, All Bond Universal). All specimens were stored in distilled water at $37^{\circ}C$ for 24 hours. Shear bond strength testing was performed with a universal testing machine at a cross head speed of 1 m/min. Data (MPa) were analyzed using one-way ANOVA and the post hoc Tukey's multiple comparison test (${\alpha}$=.05). Results: There were significant differences between Single Bond Universal, All Bond Universal, Metal Primer II and Alloy Primer, MKZ Primer, Metal & Zirconia Primer (P<.001). Conclusion: Universal Adhesive system groups indicated high shear bond strength value bonded to Ni-Cr alloy than that of conventional system groups using primers except Metal Primer II. Within the limitations of this study, improvement of universal adhesive systems which can be applied to all types of restorations is recommended especially non-precious metal alloy. More research is needed to evaluate the effect of silane inclusion or exclusion in universal adhesive systems.

Development of a Novel Long-Range 16S rRNA Universal Primer Set for Metagenomic Analysis of Gastrointestinal Microbiota in Newborn Infants

  • Ku, Hye-Jin;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.812-822
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    • 2014
  • Metagenomic analysis of the human intestinal microbiota has extended our understanding of the role of these bacteria in improving human intestinal health; however, a number of reports have shown that current total fecal DNA extraction methods and 16S rRNA universal primer sets could affect the species coverage and resolution of these analyses. Here, we improved the extraction method for total DNA from human fecal samples by optimization of the lysis buffer, boiling time (10 min), and bead-beating time (0 min). In addition, we developed a new long-range 16S rRNA universal PCR primer set targeting the V6 to V9 regions with a 580 bp DNA product length. This new 16S rRNA primer set was evaluated by comparison with two previously developed 16S rRNA universal primer sets and showed high species coverage and resolution. The optimized total fecal DNA extraction method and newly designed long-range 16S rRNA universal primer set will be useful for the highly accurate metagenomic analysis of adult and infant intestinal microbiota with minimization of any bias.

Application of Environmental DNA for Monitoring of Freshwater Fish in Korea (환경유전자의 국내 담수어류 모니터링 적용 연구)

  • Kim, Jeong-Hui;Jo, Hyunbin;Chang, Min-Ho;Woo, Seung-Hyun;Cho, Youngho;Yoon, Ju-Duk
    • Korean Journal of Ecology and Environment
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    • v.53 no.1
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    • pp.63-72
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    • 2020
  • In this study, to discuss on the applicability of eDNA as a new method to investigate fish diversity at streams, we applied eDNA at 4 streams (Geum River, Ji Stream, Hwangji Stream, Seomjin River), where endangered species are inhabits, with conventional survey (cast net and kick net). The average (±standard deviation) number of species investigated by eDNA were 19 species (±4.4), and it was relatively higher than average of conventional survey, 10 species (±4.8). Most of case, in this study, eDNA was more efficient than conventional survey. However, there were errors on species identification of Korean endemic species and aliied species from eDNA, and it seems the universal primer (MiFish primer set) is not suitable for them. Furthermore, some of endangered species, caught by conventional method, was not detected by eDNA. As the present universal primer is not suitable for identify the every freshwater fish species in Korea, the complementing or development of universal primer is needed, and the eDNA application after species specific marker development for detecting specific species like endangered species should be considered. In conclusion, if the manual for field survey method by eDNA is developed, we expect applicability enlargement for water ecosystem survey.

Development of Broad-range and Specific 16S rRNA PCR for Use in Routine Diagnostic Clinical Microbiology (임상미생물 검출을 위한 광대한 범위와 특이도를 가지는 16S rRNA PCR법 개발)

  • Kim, Hyun-Chul;Kim, Yun-Tae;Kim, Hyogyeong;Lee, Sanghoo;Lee, Kyoung-Ryul;Kim, Young-Jin
    • Journal of Life Science
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    • v.24 no.4
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    • pp.361-369
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    • 2014
  • Broad-range and specific 16S rRNA gene PCR is used for detection and identification of bacterial pathogens in clinical specimens from patients with a high suspicion for infection. We describe the development of a broad-range and specific PCR primer, based on bacterial 16S rRNA, for use in routine diagnostic clinical microbiology services. The primers were designed by using conservative regions of 16S rRNA sequences from 10 strains. Ninety-eight clinical strains were isolated from clinical patient specimens. A total of 98 strains of bacteria were identified by phenotypic methods; PCR with newly designed primers and universal primers. All purified PCR products were sequenced using both forward and reverse primers on an automated DNA analyzer. In this study, we evaluated the usefulness of the newly designed primers and the universal primers for the detection of bacteria, and both these techniques were compared with phenotypic methods for bacteria detection. When we also tested 98 strains of clinical isolates with newly designed primers, about 778 bp DNA fragments were amplified and identified from all strains. Of the 98 strains, 94 strains (95.9%) correspond in comparison with phenotypic methods. The newly designed primers showed that the identities of 98 (100%) strains were the same as those obtained by universal PCR primers. The overall agreement between the newly designed primers and universal primers was 100%. The primer set was designed for rapid, accurate, and cheap identification of bacterial pathogens. We think the newly designed primer set is useful for the identification of pathogenic bacteria.

Specific Primer for Detection of Jujube Witches' Broom Phytoplasma Group (16SrV) in Korea

  • Han, Sang-Sub
    • The Plant Pathology Journal
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    • v.21 no.1
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    • pp.55-58
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    • 2005
  • In order to diagnose and differentiate jujube witches' broom (JWB) phytoplasma rapidly, oligonucleotide primer pair, 16Sr(V) F/R, for polymerase chain reactions (PCRs) was designed on the basis of 16S rRNA sequences of JWB phytoplasma. The PCR employing phytoplasma universal primer pair P1/P7 consistently amplified DNA in all tested phytoplasma isolates. But no phytoplasma DNA was detected from healthy jujube seedlings. The nested PCR, the primer pair 16S(V) F/R, about 460 bp fragment, amplified DNA in all tested JWB and related phytoplasmas including ligustrum witches' broom phytoplasma of the 16S rRNA group V, but no DNA amplification was detected from other phytoplasma strains such as groups 16SrI (Aster yellows) and 16SrXII (Stolbur group) in which mulberry dwarf phytoplasma and chrysanthemum witches' broom phytoplasma belong to, respectively. The same results were obtained from both Korean and Chinese isolates of JWB phytoplasma. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16SrV group-specific primer pair 16S(V) F/R could detect group V phytoplasmas rapidly and easily, in particular JWB phytoplasma.

The effect of silane and universal adhesives on the micro-shear bond strength of current resin-matrix ceramics

  • Sarahneh, Omar;Gunal-Abduljalil, Burcu
    • The Journal of Advanced Prosthodontics
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    • v.13 no.5
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    • pp.292-303
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    • 2021
  • PURPOSE. The aim of this in vitro study was to evaluate the effect of silane and universal adhesive applications on the micro-shear bond strength (µSBS) of different resin-matrix ceramics (RMCs). MATERIALS AND METHODS. A total of 120 slides (14 × 12 × 1 mm) were produced from 5 different RMC materials (GC Cerasmart [GC]; Brilliant Crios [BC]; Grandio blocs [GB]; Katana Avencia [KA]; and KZR-CAD HR 2 [KZR]) and sandblasted using 50 ㎛ Al2O3 particles. Each RMC material was divided into six groups according to the surface conditioning (SC) method as follows: control (G1), silane primer (G2), silane-free universal adhesive (G3), silane-containing universal adhesive (G4), silane primer and silane-free universal adhesive (G5), and silane primer and silane-containing universal adhesive (G6). Three cylindric specimens made from resin cement (Bifix QM) were polymerized over the treated surface of each slide (n = 12). After thermal cycling (10000 cycles, 5 - 55℃), µSBS test was performed and failure types were evaluated using a stereomicroscope. Data were analyzed using 2-way ANOVA and Tukey tests (α = .05). RESULTS. µSBS values of specimens were significantly affected by the RMC type and SC protocols (P < .001) except the interaction (P = .119). Except for G2, all SC protocols showed a significant increase in µSBS values (P < .05). For all RMCs, the highest µSBS values were obtained in G4 and G6 groups. CONCLUSION. Only silane application did not affect the µSBS values regardless of the RMC type. Moreover, the application of a separate silane in addition to the universal adhesives did not improve the µSBS values. Silane-containing universal adhesive was found to be the best conditioning method for RMCs.

Comparison of bond strengths of ceramic brackets bonded to zirconia surfaces using different zirconia primers and a universal adhesive

  • Lee, Ji-Yeon;Ahn, Jaechan;An, Sang In;Park, Jeong-won
    • Restorative Dentistry and Endodontics
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    • v.43 no.1
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    • pp.7.1-7.7
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    • 2018
  • Objectives: The aim of this study is to compare the shear bond strengths of ceramic brackets bonded to zirconia surfaces using different zirconia primers and universal adhesive. Materials and Methods: Fifty zirconia blocks ($15{\times}15{\times}10mm$, Zpex, Tosoh Corporation) were polished with 1,000 grit sand paper and air-abraded with $50{\mu}m$ $Al_2O_3$ for 10 seconds (40 psi). They were divided into 5 groups: control (CO), Metal/Zirconia primer (MZ, Ivoclar Vivadent), Z-PRIME Plus (ZP, Bisco), Zirconia Liner (ZL, Sun Medical), and Scotchbond Universal adhesive (SU, 3M ESPE). Transbond XT Primer (used for CO, MZ, ZP, and ZL) and Transbond XT Paste was used for bracket bonding (Gemini clear ceramic brackets, 3M Unitek). After 24 hours at $37^{\circ}C$ storage, specimens underwent 2,000 thermocycles, and then, shear bond strengths were measured (1 mm/min). An adhesive remnant index (ARI) score was calculated. The data were analyzed using one-way analysis of variance and the Bonferroni test (p = 0.05). Results: Surface treatment with primers resulted in increased shear bond strength. The SU group showed the highest shear bond strength followed by the ZP, ZL, MZ, and CO groups, in that order. The median ARI scores were as follows: CO = 0, MZ = 0, ZP = 0, ZL = 0, and SU = 3 (p < 0.05). Conclusions: Within this experiment, zirconia primer can increase the shear bond strength of bracket bonding. The highest shear bond strength is observed in SU group, even when no primer is used.

Phytoplasma specific primer for detection of jujube witches′ broom group(16SrV) in Korea and China

  • Sangsub Han;Lee, Sanghun;Mengjun Liu;Byeongjin Cha
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.136.2-137
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    • 2003
  • In order to diagnose and differentiate jujube witches' broom (JWB) phytoplasma rapidly, oligonucleotide primer pair, 16Sr(V) F/R, for polymerase chain reactions (PCRs) was designed on the basis of 165 rRNA sequences of JWB phytoplasma. The PCR employing phytoplasma universal primer pair P1/P7 consistently amplified DNA in all tested phytoplasma isolates. But no phytoplasma DNA was detected in healthy jujube seedlings. The nested PCR, the primer pair 16S(V) F/R, about 460 bp fragment, amplified DNA in all tested JWB and related phytoplasmas including LiWB phytoplasma of the 165 rRNA group V, but no DNA amplification was detected from other phytoplasma strains such as group 16SrI (Aster yellows) and group 16SrⅩII (Stolbur group) phytoplasmas in which mulberry dwarf phytoplasma and chrysanthemum witches broom phytoplasma are belonged to, respectively The same results were obtained from both Korean- and Chinese-isolates of JWB. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16S rRNA group V specific primer pair 16S(V) F/R could detect group V phytoplasma rapidly and easily, in particular JWB phytoplasma.

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PCR Based Detection of Phellinus linteus using Specific Primers Generated from Universal Rice Primer(URP) Derived PCR Polymorphic Band

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;Lee, Byoung-Moo;Cho, Soo-Muk;Kim, Ki-Tae;Seo, Geon-Sik;Go, Seung-Joo
    • Mycobiology
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    • v.30 no.4
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    • pp.202-207
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    • 2002
  • This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.

Identification of Phellinus linteus by Morphological Characteristics and Molecular Analysis (형태적.분자생물학적 방법에 의한 Phellinus linteus의 동정에 관한 연구)

  • Kim, Sang-Hee;Kim, Soo-Ho;Sung, Jae-Mo;Harrington, Thomas C.
    • The Korean Journal of Mycology
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    • v.27 no.5 s.92
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    • pp.337-340
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    • 1999
  • The context and upper surface of Phellinus basidiocarp become blackened, rimose and woody. The basidiocarp is sessile, dimidiate and elongate. The basidiospores are pigmented and ovoid to globose. Hymenial setae are $17{\sim}35{\times}6{\sim}8{\mu}m$. Nineteen isolates of Phellinus species, including Phellinus linteus, were used for sequencing of the internal transcribed spacer (ITS) region of the nuclear rDNA. Based on these sequence data, specific primers were designed for identification of Phellinus linteus isolates in Korea. The specific primers were within the ITS1 and ITS2 regions and were nested within the universal primers flanking the spacer regions. A total of four primers (the universal primers ITS-1F and ITS-4, and the specific primers PL-F and PL-R) were used for detection of Phellinus linteus collected in Korea. The length of the four amplification products of Phellinus linteus DNA were 800 bp (ITS-1F/ITS-4), two bands of about 720 bp (ITS-1F/PL-R and PL-F/ITS-4), and 610 bp (PL-F/PL-R). Among 23 isolates of Phellinus species collected in Korea, Thirteen isolates were identified as Phellinus linteus based on the presence of the four bands. The other species produced only the single ITS-1F/ITS-4 product.

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