• International Journal of Oral Biology
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• 제46권3호
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• pp.140-145
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• 2021

This study aimed to develop Lautropia mirabilis-specific quantitative real-time polymerase chain reaction (qPCR) primers based on the sequence of DNA-directed RNA polymerase subunit beta gene. The PrimerSelect program was used in designing of the qPCR primers, RTLam-F4 and RTLam-R3. The specificity of the qPCR primers were performed by conventional PCR with 37 strains of 37 oral bacterial species, including L. mirabilis. The sensitivity of the primers was determined by qPCR with the serial dilution of purified genomic DNA of L. mirabilis KCOM 3484, ranged from 4 ng to 4 fg. The data showed that the qPCR primers could detect only L. mirabilis strains and as little as 40 fg of genome DNA of L. mirabilis KCOM 3484. These results indicate that this qPCR primer pair (RTLam-F4/RTLam-R3) may be useful for species-specific detection of L. mirabilis in epidemiological studies of oral bacterial infectious diseases such as periodontal disease.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권4호
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• pp.213-220
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• 2003

Polymerase chain reaction (PCR) is one of the most widely used analytical tool and is an important module that would benefit from being miniaturized and integrated onto diagnostic or analytical chips. There are potentially two different approaches for the miniaturization of the PCR module: chamber-type and flow-type micro-PCR. These miniaturized PCRs have distinct characteristics and advantages. In this article, we review the necessity of micro-PCR, the materials for the chip fabrication, the surface modification, and characteristics of the two types of micro-PCR. The motivation underlying the development of micro-PCR, the advantages and disadvantages of the various materials used in fabrication and the surface modification methods will be discussed. And finally, the precise features of the two different types of micro-PCR will be compared.

• 한국환경보건학회지
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• 제28권2호
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• pp.193-202
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• 2002

The polymerase chain reaction(PCR) of target lacZ and uidA genes were used to detect total coliform and Escherichia coli for determining water quality, respectively. Of 109 spring waters, coliform were detected from 38 spring waters by lacZ PCR method but 21 spring waters by culture method accepted by the Ministry of Environment for water quality monitoring. The lacz PCR method gave the results statistically equivalent to those of the culture method(kappa=0.62, McNemar=17.00). The uidA PCR method gave the same results to those of the culture method. The sensitivity and specificity of coliform and E. coli by PCR method were 100% and 80.7%, respectively. Therefore, PCR can be used for the rapid identification of Escherichia coli and coliform in potable water using uidA and lacZ.

• 한국동물생명공학회지
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• 제34권4호
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• pp.259-266
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• 2019

Emulsion polymerase chain reaction (PCR) is performed on compartmentalized DNA, allowing a large number of PCR reactions to be carried out in parallel. Emulsion PCR has unique advantages in DNA amplification. It can be applied in many molecular biological assays, especially those requiring highly sensitive and specific DNA amplification. This review discusses the principle of emulsion PCR and its applications in biotechnology. Related technologies are also discussed.

• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.881-884
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• 2000

A polymerase chain reaction (PCR) was performed to rapidly identify Lactobacillus plantarum from type strains and kimchi samples. The PCR experiments were carried out using specific oligonucleotide primer sets based on the 16S rRNA gene sequences of L. plantarum. The expected DNA amplificate of 419 bp was obtained when either purified DNA or whole cells of L. plantarum strains reacted with LP primers, yet not with any of the other strains. The PCR product was confirmed by DNA sequencing. Accordingly, since the PCR method used is simple, specific, and rapid, it will be useful for monitoring and evaluation L. plantarum in the mixed microbial population found in kimchi.

• 한국식물병리학회지
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• 제12권2호
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• pp.191-196
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• 1996

위축, 황화, 총생 증상 등 전형적인 병징을 나타내는 phytoplasma에 감염된 식물에서 phytoplasma만을 특이적으로 검출하기 위하여 polymerase chain reaction(PCR) 방법을 이용하였다. Phytoplasma의 16S rRNA gence의 DNA 단편을 증폭하기 위하여 1.4 kb primers (forward, 5` -GTTGATCCTGGCTCAGGATT-3` 와 reverse, 5` -AACCCCGAGAACGTATTCACC -3`)를 사용하여 증폭한 결과, phytoplasma에 이병된 오동나무, 라일락 및 미역취에서는 약 1.4 kbp의 위치에서 특이 band가 검출되었으나 control로 사용한 건전주에서는 어떠한 band 검출되지 않았다. 위의 결과를 재확인 하기 위하여 약 0.5 kb의 primers(forward, 5` -ACGAAAGCGTGGGGAGCAAA-3` 와 reverse, 5` -GAAGTCGAGTTGCAGACTTC-3`)를 사용하여 증폭한 결과, 0.5 kb의 위치에서 특이 band가 검출되었으나 control로 사용한 건전주에서는 어떠한 band도 검출되지 않았다. Phytoplasma에 이병된 식물의 PCR 반응산물을 제한효소인 AluI으로 처리하 sruf과, 오동나무와 라일락에서는 동일한 band pattern을 나타내어 서로 유연관계가 가까운 phytoplasma인 것으로 생각되며, 미역취에서는 이들과는 다른 band pattern을 나타내어 오동나무와 라일락의 phytoplasma와는 유연관계가 먼 것으로 추측된다.

• 미생물학회지
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• 제40권3호
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• pp.254-256
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• 2004

Polymerase chain reaction (PCR) is a powerful tool for precisely amplifying selected DNA sequences that have had a broad impact on genomic studies. When examining human $\alpha$- and $\beta$- tryptase genes which have 95% DNA homology, inconsistent PCR amplification of genomic sequences hampered our progress. This study suggests that long PCR technique on the original DNA digested with restriction enzymes improves both efficiency and sensitivity of PCR. These improved results seem to derived from the effective denaturation of the original genomic DNA template or reduction of formation of secondary structures that block either primer annealing or extension in PCR. Elimination of homo- or hetero-duplex products derived from highly homologous genes provides an additional advantage in this study. This communication describes how the use of restriction enzymes improved these efficiencies, and also facilitated studies of highly homologous genes including tryptase genes.

• 한국동물위생학회지
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• 제23권3호
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• pp.227-233
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• 2000

Salmonella enteritidis is the most prevalent etiologic agents of foodborne acute gastroenteritis. Direct isolation and identification of S enteritidis are time consuming work and not so highly sensitive. This study was conducted to develop for the specific detection of S enteritidis using polymerase chain reaction(PCR). PCR primers were selected to amplify a 351-base pair(bp) DNA fragment from the salmonella plasmid virulence A(spv A) gene of S enteritidis. With the primers, 351 bp DNA products were amplified from S enteritidis but not from other B, D, Cl serogroup Salmonella spp. It was sensitive to detect up to 40 pg of template DNA by agarose gel electrophoresis. This PCR assay is very rapid and specific method and less time consuming than the standard bacteriological methods.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2016년도 제50회 동계 정기학술대회 초록집
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• pp.375.1-375.1
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• 2016

Polymerase chain reaction (PCR) has revolutionized genetics and become one of the most popular techniques in modern biological and medical sciences. It can be used not only as an in vitro DNA amplification method but also used in many bioassay applications. The PCR can be used to exponentially produce a large number of DNA copies from a small quantity of DNA molecules in a few hours. However, as unwanted DNA fragments are also often manufactured, the amplification efficiency of PCR is decreased. To overcome this limitation, several nanomaterials have been employed to increase the specificity of the PCR reaction. Recently, graphene has attracted a great interest for its excellent electron transfer, thermal and biocompatibility. Especially, gold nanoparticle-coated graphene oxide (GO/AuNPs) led to enhance electron and thermal transfer rate and low-charge transfer resistance. Therefore, we report the development of a demonstration for the PCR efficiency using a large-scale production of the GO and combination of gold nanoparticles. Because a thermal conductivity is an important factor for improving the PCR efficiency in different DNA polymerases and different size samples. When PCR use GO/AuNPs, the result of transmission electron microscopy and real-time quantitative PCR (qPCR) showed an enhanced PCR efficiency. We have demonstrated that GO/AuNPs would be simply outperformed for enhancing the specificity and efficiency of DNA amplification procedure.

• 대한수의학회지
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• 제39권1호
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• pp.90-95
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• 1999

A species-specific 760 base pair(bp) BamHI to EcoRI DNA fragment(fMG-2) of lipoprotein gene was isolated from a Mycoplasma gallisepticum(M gallisepticum) genomic library. Based on the DNA sequence data of fMG-2, a pair of 25bp primers was synthesized. When used in the polymerase chain reaction(PCR), 732bp DNA products were amplified from 6 standard strains and 10 field isolates of M gallisepticum, but not from 2 Mycoplasma synoviae and 7 other Mycoplasma species. The lower detection limit was 100fg of the genomic DNA. Identity of the PCR products was confirmed by comparison of patterns of restriction endonuclease analysis with AseI, DraI, EcoRV and SspI.