• Title/Summary/Keyword: 프라이머

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Identification of Pleurotus ostreatus cultivars with the application of multiplex-simple sequence repeat markers (Multiplex SSR마커를 이용한 느타리(Pleurotus ostreatus) 품종 판별)

  • Choi, Jong In;Jung, Hwa Jin;Na, Kyeong sook;Oh, Min-Ji;Kim, Min-Keun;Ryu, Jae-San
    • Journal of Mushroom
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    • v.19 no.1
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    • pp.76-80
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    • 2021
  • To develop a method for the differentiation of Pleurotus ostratus cultivars, the multiplex-simple sequence repeat (SSR) primer set based on the SSRs obtained from whole genomic DNA sequence analysis was designed with two polymerase chain reaction (PCR) primer sets. These SSR primer sets were employed to distinguish 10 cultivars and strains. Twenty polymorphic markers were selected based on the genotyping results. PCR with each primer produced 1-4 distinct bands ranging in size from 150 to 350 bp, which was within the expected range. However, since a sole SSR marker was unable to detect polymorphisms in every cultivar, multiplex PCRs with composite PCR primer sets were employed. The multiplex primer, "166+115," completely discriminated 12 cultivars and strains with 40 loci, which were 12 more than the simple arithmetic addition of each locus of the primers 115 and 166. These results might be useful to provide an efficient method for the differentiation of P. ostreatus cultivars with separate PCRs for the quality control of spawn and protection of breeders' rights.

Development of Raw Material Identification Method of Changnan-jeot and Gaiyang-jeot Using Multiplex PCR and Real-Time PCR (Multiplex PCR과 Real-Time PCR을 이용한 창난젓과 가이양젓 원료 검사법 개발)

  • Choi, Seong Seok;Seo, Yong Bae;Kim, Jong-Oh;Yang, Ji-Young;Shin, Jiyoung;Kim, Gun-Do
    • Journal of Food Hygiene and Safety
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    • v.36 no.4
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    • pp.289-297
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    • 2021
  • In this study, multiplex PCR and real-time PCR were performed on Theragra chalcogramma (walleye pollock), Pangasianodon hypophthalmus (iridescent shark) and their processed foods, such as changnan-jeot and gaiyang-jeot (salted iridescent shark intestine). Species-specific primers for T. chalcogramma and P. hypophthalmus were designed, and genomic DNA was directly extracted from each sample to perform single PCR and multiplex PCR. As a result of PCR, in the case of single PCR, PCR bands of T. chalcogramma (297 bp) and P. hypophthalmus (132 bp) were identified, and in the case of multiplex PCR, it was confirmed that amplification occurred without cross-reaction between T. chalcogramma and P. hypophthalmus. As a result of checking the PCR sensitivity, the concentration of genomic DNA was detected up to 0.1 ng/µL in both single PCR and multiplex PCR. The real-time PCR results showed that the average Ct value of T. chalcogramma was 20.765±0.691, and the average Ct value of P. hypophthalmus sample was 35.719±1.828 in the T. chalcogramma species-specific primers. In the P. hypophthalmus species-specific primers, the average Ct value of the T. chalcogramma sample was 35.996±1.423, and the mean Ct value of the P. hypophthalmus sample was 20.096±0.793. These results demonstrated the significant differences in the efficiency, specificity and cross-reactivity of species-specific primers in real-time PCR. Based on these findings, 7 of changnan-jeot or gaiyang-jeot products were confirmed by multiplex PCR and real-time PCR, and valid results were confirmed in all samples.

Species-specific Marker Development for Environmental DNA Assay of Endangered Bull-head Torrent Catfish, Liobagrus obesus (멸종위기어류 퉁사리의 환경 DNA 분석을 위한 종 특이 마커 개발)

  • Yun, Bong Han;Kim, Yong Hwi;Sung, Mu Sung;Han, Ho-Seop;Han, Jeong-Ho;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.34 no.3
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    • pp.208-217
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    • 2022
  • We wanted to develop a real-time PCR assay capable of detecting Liobagrus obesus in environmental DNA (eDNA) extracted from freshwater samples using a pair of species-specific primers and probe for the endangered fish, L. obesus. The species-specific primers and probe were designed in consideration of single nucleotide polymorphisms between 65 species of freshwater fish living in the Republic of Korea within the cytochrome b (cytb) gene of mitochondrial DNA. The species-specific primers and probe, in the real-time PCR assay, showed high specificity as only the L. obesus genomic DNA (gDNA) was found to be positive in the specificity verification using 65 species gDNA of freshwater fish in the Republic of Korea. In addition, in the detection limit analysis using the serial dilution concentrations of L. obesus gDNA, it was found that it was possible to detect up to 0.2 pg, showing high sensitivity. Afterwards, using the species-specific primers and probe, real-time PCR assay was performed on freshwater samples obtained from 8 stations in the mid-upper basin of Geum River. As a result, the cytb gene of L. obesus was detected in total 5 stations including all 3 stations where this species was collected at the time of field survey. Therefore, the species-specific primers and probe developed in present study, and the real-time PCR assay using them, can accurately detect the cytb gene of L. obesus from eDNA samples, which can be utilized to monitor the existing habitats of this species and to discover potential new habitats.

In vitro cytotoxicity of self-etching primers (자가 산부식 프라이머의 세포독성에 관한 실험적 연구)

  • Rhee, Chang-Hoon;Kim, In-Ryeon;Kim, Gyoo-Cheon;Kim, Seong-Sik;Son, Woo-Sung
    • The korean journal of orthodontics
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    • v.36 no.6
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    • pp.422-433
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    • 2006
  • Objective: Several ions and components are released from self-etching primers in the oral cavity. This may cause injury to the periodontal tissues throughout orthodontic treatment. The purpose of this study was to assess the cytotoxicity of self-etching primers to HGF-1, HaCaT, and RHEK cells. Method: Transbond XT Primer (3M Unitek, Monrovia, CA, USA), and self-etching primers, Clearfil SE Bond (Kuraray, Osaka, Japan), Transbond Plus SEP (3M Unitek, Monrovia, CA, USA), and Adper Prompt L-Pop (3M Unitek, Monrovia, CA, USA), were evaluated by MTT assay, and cellular changes were also observed. Results: In all cells after 72 hours with all primers, severe morphological changes such as atrophy and necrosis were observed. In the MTT assay using HGF-1, Clearfil SE Bond, Transbond XT Primer, Transbond Plus SEP, and Adper Prompt L-Pop were lined up in order of ascending cytotoxicity When using HaCaT, Clearfil SE Bond, Adper Prompt L-Pop, Transbond Plus SEP, and Transbond XT Primer were lined up in order of ascending cytotoxicity. When using RHEK, Clearfil SE Bond, Transbond XT Primer, Adper Prompt L-Pop, and Transbond Plus SEP were lined up in order of ascending cytotoxicity. Conclusion: The result of this study shows that care is needed because self-etching primers show cytotoxic properties similar to conventional primers.

Effects of primers on the microtensile bond strength of resin cements to cobalt-chromium alloy (레진 시멘트와 코발트 크롬 합금의 미세인장결합강도에 다양한 프라이머들이 미치는 영향)

  • Jung, Hong-Taek;Campana, Shiela A.;Park, Jin-Hong;Shin, Joo-Hee;Lee, Jeong-Yol
    • The Journal of Korean Academy of Prosthodontics
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    • v.57 no.2
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    • pp.95-101
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    • 2019
  • Purpose: The aim of this study is to evaluate the effects of various primers on the microtensile bond strength (${\mu}TBS$) of resin cements to cobalt-chromium (Co-Cr) dental casting alloy. Materials and methods: Four adhesive primers (Universal primer, Metal primer II, Alloy primer, and Metal/Zirconia primer) and two resin cements (Panavia F2.0, G-CEM LinkAce) were tested. One hundred fifty Co-Cr beams were prepared from Co-Cr ingots via casting ($6mm\;ength{\times}1mm\;width{\times}1mm\;thick$). The metal beams were randomly divided into ten groups according to the adhesive primers and resin cements used; the no-primer groups served as the control (n = 15). After sandblasting with aluminum oxide ($125{\mu}m$ grain), the metal and resin cements were bonded together using a silicone mold. Prior to testing, all metal-resin beams were examined under stereomicroscope, and subjected to the ${\mu}TBS$ test. The mean value of each group was analyzed via one-way ANOVA with Tukey's test as post hoc (${\alpha}=.05$) using SPSS software. Results: The mean ${\mu}TBS$ of all groups was ranged from 20 to 28 MPa. There is no statistically significant difference between groups (P > .05). Mixed failure, which is the combination of adhesive and cohesive failures, is the most prevalent failure mode in both the Panavia F2.0 and G-Cem LinkAce groups. Conclusion: The ${\mu}TBS$ of all tested groups are relatively high; however, the primers used in this study result in no favorable effect in the ${\mu}TBS$ of Panavia F2.0 and G-Cem LinkAce resin cement to Co-Cr alloy.

Effect of various surface treatment methods of highly translucent zirconia on the shear bond strength with resin cement (고투명도 지르코니아의 다양한 표면처리 방법이 레진시멘트와의 전단결합강도에 미치는 영향)

  • Yu-Seong Kim;Jin-Woo Choi;Hee-Kyung Kim
    • The Journal of Korean Academy of Prosthodontics
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    • v.61 no.3
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    • pp.179-188
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    • 2023
  • Purpose. The purpose of this study was to evaluate the effect of surface treatments on the shear bond strength of two types of zirconia (3-TZP and 5Y-PSZ) with resin cement. Materials and methods. Two different types of zirconia specimens with a fully sintered size of 14.0×14.0×2.0 mm3 were prepared, polished with 400, 600, and 800 grit silicon carbide paper, and buried in epoxy resin. They were classified into four groups each control, sandblasting, primer, and sandblasting & primer. Cylindrical resin adhered to the surface-treated zirconia with resin cement. It was stored in distilled water (37℃) for 24 hours, and a shear bond strength test was performed. The normality of the experimental group was confirmed with the Kolmogorov-Smirnov & Shapiro-Wilk test. The interaction and statistical difference were analyzed using a two-way ANOVA. A post-hoc analysis was performed using Dunnett T3. Results. As a result of two-way ANOVA, there was no significant difference in shear bonding strength between zirconia types (P > .05), but there was a significant correlation in the sandblasting, primer, and alumina sandblasting & primer group (P < .05). Dunnett T3 post-test showed that, regardless of the type of zirconia, shear bonding strength was sandblasting & primer > Primer > sandblasting > control group (P < .05). Conclusion. There was no difference in shear bond strength between the types of zirconia. The highest shear bond strength was shown when the mechanical and chemical treatments of the zirconia surface was performed simultaneously.

PCR 방법에 의한 세균성 식중독균의 다중${\cdot}$동시 검출

  • Park, Ju-Yeon;Mun, Seon-Yeong;Lee, Gyeong-Ha;Nam, Jeong-Ok;Nam, Bo-Ra;Kim, Jin-Man
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2005.10a
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    • pp.346-349
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    • 2005
  • 병원성미생물을 분리${\cdot}$동정하는 방법으로 선택배지를 이용하는 검사방법은 4${\sim}$7일의 시간이 소요되는 단점이 있으며, 기존의 PCR방법은 대부분 단일 병원성 미생물만을 검출할 수 있는 반면 본 연구에서 사용된 시스템은 6종류의 세균성 식중독균을 한 번의 분석으로 확인할 수 있었기 때문에 비용과 분석시간을 대폭 절감하는 효과가 있었다. Fig. 1은 6종류의 세균성 식중독균이 혼합된 시료에 각각의 단일 프라이머를 이용하여 특이성을 확인한 결과이며, Fig. 2는 멀티플렉스 프라이머를 이용하여 각각의 세균성 식중독균 열 추출 시료에 대한 특이성을 확인한 결과이다. PCR 산물들이 사다리(ladder) 형태로 증폭됨으로써 동시에 6종류의 세균성 식중독균을 용이하게 확인할 수 있었다(Lane 1${\sim}$Lane 6). 또한 3종류의 세균성 식중독균이 혼합된 열 추출시료에서도 특이적인 PCR 증폭 산물들을 탐지할 수 있었다(Lane 10 & Lane 11).

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Comparison of Adhesion Performance According to Concrete Moisture Content of Primer Mixed with Cement and Modified Polymer (시멘트와 변성 폴리머를 혼합한 프라이머의 콘크리트 함수율에 따른 부착성능 비교)

  • Kang, Hyo-Jin;An, Ki-Won;Oh, Sang-Keun
    • Proceedings of the Korean Institute of Building Construction Conference
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    • 2019.11a
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    • pp.231-232
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    • 2019
  • In this study, primers were prepared by mixing cement and modified polymer, and the adhesion to the substrate surface was enhanced by using cement which is the same material as the basement outer wall. The improved primer is used to verify the adhesion performance of the substrate in wet concrete environments.

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Development of Duplex PCR Method for Simultaneous Detection of Rabbit (Oryctolagus cuniculus) and Cat (Felis catus) Meats (Duplex PCR을 이용한 토끼(Oryctolagus cuniculus)와 고양이(Felis catus) 육류의 동시 검출법 개발)

  • Hong, Yeun;Kim, Mi-Ju;Yang, Seung-Min;Yoo, In-Suk;Kim, Hae-Yeong
    • Journal of Applied Biological Chemistry
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    • v.58 no.4
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    • pp.383-387
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    • 2015
  • A duplex polymerase chain reaction (PCR) detection method was developed to authenticate the use of cat and rabbit in food and to prevent unlawful distribution of illegally butchered meat in both domestic and imported food market. Species-specific primers were designed targeting mitochondrial cytochrome b gene. The sizes of PCR products were 191 bp for cat and 101 bp for rabbit, which were relatively small for better application of the detection method on processed foods. Specificities of primers were verified using 21 animal species including cat and rabbit. Limit of detection was examined by serial dilution of the sample DNA and confirmed as 0.005 ng for rabbit and 0.0005 ng for cat using Bioanalyzer. The developed duplex PCR method showed specificity and sensitivity in the identification of two target species.