• Title/Summary/Keyword: Specific DNA

Search Result 2,805, Processing Time 0.035 seconds

Utilization of DNA Marker-Assisted Selection in Korean Native Animals

  • Yeo, Jong-sou;Kim, Jae-Woo;Chang, Tea-Kyung;Pake, Young-Ae;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.5 no.2
    • /
    • pp.71-78
    • /
    • 2000
  • The recent progress od DNA technologies including DNA fingerprinting (DFP) and random amplified DNA polymorphism (RAPD) analysis make it possible to identify the specific genetic trits of animals and to analyze the genetic diversity and relatedness between or withinspecies or populations. Using those techniquse, some efforts to identify and develop the specific DNA markers based on DNA polymorphism, which are related with economic traits for Korean native animals, Hanwoo(Korean native cattle),Korean native pig and Korean native chicken, have been made in Korea for recent a few years. The developed specific DNA markers successfully characterize the Korean native animals as the unique Korean genetic sources, distinctively from other imported breeds. Some of these DNA markers have been related to some important economic traits for domestic animals, for example, growth rate and marbling for Honwoo, growth rate and back fat thinkness fornative pig, and growth rate, agg weight and agg productivity for native chicken. This means that those markers can be used in important marker-assised selection (MAS) of Korean native domestic animals and further contribute to genetically improve and breed them.

  • PDF

Extension of a 5'- or 3'-end Genomic DNA Sequence by a Single PCR Amplification

  • Jeon, Taeck J.
    • Journal of Integrative Natural Science
    • /
    • v.1 no.3
    • /
    • pp.230-233
    • /
    • 2008
  • A simple and rapid method is described for extending the 5'- or 3'-end genomic sequence of a known partial sequence by only a single round of PCR. This method involves digesting and ligating genomic and plasmid DNAs, and amplifying the 5'-upstream or 3'-end downstream sequence of the known DNA sequence, using two primers, one gene specific and the other plasmid specific. A single round of PCR amplification is sufficient to produce gene-specific bands detectable in gels. By using this approach, 5'-end genomic sequence of the D-amoeba sams gene was extended.

  • PDF

Development of a Method for Rapid Analysis of DNA Hybridization (측방유동방식 신속 DNA 교잡 분석법의 개발)

  • 정동석;최의열
    • Korean Journal of Microbiology
    • /
    • v.39 no.2
    • /
    • pp.114-117
    • /
    • 2003
  • In molecular biology, it is necessary to develop an easy and rapid method to identify a specific DNA sequence. Though Southern and Northern blot techniques have been used widely for the analysis of gene structure and function, those methods are inconvenient in the points that we need to control incubation temperature, time, and other parameters to get the final result. In this study, we report a new method for the rapid analysis of specific DNA sequence with the modification of an immunochromatographic method. The lateral flow DNA analysis strip is composed of a sample pad, a nitrocellulose membrane for the separation and propagation of analytes, and an absorption pad for the generation of capillary action. Capture DNA was immobilized on the membrane by UV cross-linking and target DNA was labeled with Cy-5 for signaling. The samples containing target DNA were applied onto the sample pad, incubated for 15 min for separation, and scanned with a GSI fluorescence scanner. Though the hybridization reaction occurs in a short time without any washing steps, there appears to be little cross hybridization between the different sequences. The result showed a possibility that the new method can be used for the rapid identification of specific DNA sequence among the samples.

A STUDY ON THE IDENTIFICATION OF Porphyromonas endodontalis BY PCR USING SPECIES SPECIFIC PRIMERS FOR THE 16S rDNA (16S rDNA sequence에 대한 종특이성 primer를 이용한 중합효소연쇄반응증폭에 의한 Porphyromonas endodontalis의 동정에 관한 연구)

  • Eom, Seung-Hee;Lim, Sung-Sam;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
    • /
    • v.24 no.1
    • /
    • pp.13-25
    • /
    • 1999
  • P. endodontalis which was known to be associated with the infected root canals and periapical lesions is very difficult to detect by culture methods or traditional methods. Detection of bacteria using polymerase chain reaction(PCR) for 16S ribosomal DNA(rDNA) is fast, simple, and accurate with relatively small amount of target cells. 16S rDNA consist of conserved regions those are same to all species, and variable regions which represent species specificity. The 16S rDNA sequences of P. endodontalis and P. gingivalis were aligned and two highly variable regions were selected as a pair of species specific oligonucleotide primers for P. endodontalis. And then the pair of primers for PCR amplification was synthesized to identify P. endodontalis. The sequences of the species specific primers for the 16S rDNA of P. endodontalis were as follows ; sense primer[endo1]: 5'-CTATATTCTTCTTTCTCCGCATGGAGGAGG-3' antisense primer[endo2]: 5'-GCATACCTTCGGTCTCCTCTAGCATAT-3' In this study, for the identification of P. endodontalis without culture from the mixed clinical samples, PCR was done with species specific primers for the 16S rDNA sequences of P. endodontalis. The results were as follows : 1. The species specificity of the primers for the 16S rDNA of P. endodntalis was determined by the PCR methods. About 490bp amplicon which was specific only for P. endodntalis was produced with P. endodontalis. No amplicon was produced by PCR with other strains similar to P. endodontalis. 2. The synthesized species specific primers reacted with conventionally identified P. endodontalis which we have in conservative dentistry laboratory. 3. The identification of P. endodontalis using PCR technique with samples collected from infected root canals or periapical lesions was more sensitive than that of culture methods. 4. Seven samples revealed including P. endodontalis by PCR technique. Five of them were related with pains, two of them with sinus tract, three of them with foul odor, and three of them with purulent drainage. P. endodontalis was shown to have great relation with pains.

  • PDF

Development of strain-specific polymerase chain reaction primers to detect Fusobacterium hwasookii strains

  • Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
    • /
    • v.46 no.4
    • /
    • pp.155-159
    • /
    • 2021
  • This study aimed to develop strain-specific polymerase chain reaction (PCR) primers to detect Fusobacterium hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1256, F. hwasookii KCOM 1258, and F. hwasookii KCOM 1268 on the basis of nucleotide sequences of a gene specific to each strain. The unique genes for each F. hwasookii strain were determined on the basis of their genome sequences using Roary. The strain-specific PCR primers based on each strain-specific gene were designed using PrimerSelect. The specificity of each PCR primer was determined using the genomic DNA of the 5 F. hwasookii strains and 25 strains of oral bacterial species. The detection limit and sensitivity of each strain-specific PCR primer pair were determined using the genomic DNA of each target strain. The results showed that the strain-specific PCR primers correspond to F. hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1258, F. hwasookii KCOM 1256/F. nucleatum subsp. polymorphum KCOM 1260, or F. hwasookii KCOM 1268/Fusobacterium sp. oral taxon 203 were developed. The detection limits of these strain-specific PCR primers ranged from 0.2 to 2 ng of genomic DNA for each target strain. The results suggest that these strain-specific PCR primers are valuable in quality control for detecting specific F. hwasookii strains.

Development of Genus- and Species-Specific Probe Design System for Pathogen Detection Based on 23S rDNA

  • Park Jun-Hyung;Park Hee-Kyung;Kang Byeong-Chul;Song Eun-Sil;Jang Hyun-Jung;Kim Cheol-Min
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.5
    • /
    • pp.740-747
    • /
    • 2006
  • Amplification by universal consensus sequences in pathogenic bacterial DNA would allow rapid identification of pathogenic bacteria, and amplification of genus-specific and species-specific sequences of pathogenic bacterial DNA might be used for genotyping at the genus and species levels. For design of probes for molecular diagnostics, several tools are available as stand-alone programs or as Web application. However, since most programs can design only a few probe sets at one time, they are not suitable for large-scale and automatic probes design. Therefore, for high-throughput design of specific probes in diagnostic array development, an automated design tool is necessary. Thus, we developed a Web-based automatic system for design of genus-specific and species-specific probes for pathogen detection. The system is available at http://www.miprobe.com.

Specific tail swelling pattern in hypo-osmotic solution as a predictor of DNA fragmentation status in human spermatozoa

  • Kim, Sung Woo;Nho, Eun Jee;Lee, Joong Yeup;Jee, Byung Chul
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.46 no.4
    • /
    • pp.147-151
    • /
    • 2019
  • Objective: The aim of this study was to investigate DNA fragmentation status in human spermatozoa according to specific tail swelling patterns determined via hypo-osmotic swelling test (HOST). Methods: Frozen semen samples from 21 healthy donors were thawed and prepared by the swim-up technique for use in intracytoplasmic sperm injection. The semen samples were treated for 5 minutes as part of the HOST procedure and then underwent the sperm chromatin dispersion test using a Halosperm kit. DNA fragmentation status (large halo, medium halo, small halo, no halo, or degraded) and the specific tail swelling pattern ("a"-"g") were assessed at the level of a single spermatozoon. A total of 42,000 spermatozoa were analyzed, and the percentage of spermatozoa without DNA fragmentation (as evidenced by a large or medium halo) was assessed according to the specific tail swelling patterns observed. Results: The HOST examinations showed that > 93% of spermatozoa across all types displayed no DNA fragmentation. The percentage of spermatozoa without DNA fragmentation was 100% in type "d", 98.67% in type "g", and 98.17% in type "f" spermatozoa. Conclusion: We found that the type "d" spermatozoa displayed no DNA fragmentation, but the other types of spermatozoa also displayed very low rates of DNA fragmentation. This result may be associated with the processing of the spermatozoa by density gradient centrifugation and the swim-up technique.

Gender Determination of X and Y-Specific Alphoid Repeat Sequences by PCR (PCR에 의한 X,Y-Specific Alphoid Repeat Sequences의 분석)

  • Choi, Dong-Ho;Kang, Pil-Won;Lee, Yang-Han;Han, Myun-Soo
    • Analytical Science and Technology
    • /
    • v.12 no.1
    • /
    • pp.80-83
    • /
    • 1999
  • Recently, it has been possible to the alphoid gene, which has X and Y specificity, and determine the sex from human physical evidence using PCR methods. Samples from single sources, PCR method applied to the alphoid gene results in highly sensitive and accurate results even when only 60 pg of the genomic DNA was available for sex determination. Even for samples containing DNA from more than one gender source where the female DNA was present in the amount 10 times than that of the male, sex determination was possible. Therefore, this result suggests that alphoid gene, which has X and Y specificity, could be used effectively for sex determination in case of mixed DNA samples from biological evidence.

  • PDF

A DNA Microextractor Using Crossed Field Electrophoresis (교차 전기영동법을 이용한 극소형 DNA 추출기)

  • Yi Soyeon;Seo Kyoung-Sun;Cho Young-Ho
    • Transactions of the Korean Society of Mechanical Engineers A
    • /
    • v.28 no.8 s.227
    • /
    • pp.1135-1139
    • /
    • 2004
  • This paper presents a microextractor for the separation of DNA molecules by their sizes. The DNA microextractor immobilizes the DNA molecules of specific size in the micropillar array by adjusting the period of the crossed electric field, thus providing a starting-point independent target DNA extraction method without separation process monitoring. The DNA microextractor has been fabricated by a three-mask micromachining process. The velocity of three different DNA molecules has been measured at the electric field of E=5V/0.8cm in the fabricated DNA microextractor, resulting in the reorientation times of $4.80{\pm}0.44sec,\;7.12{\pm}0.75sec$, and $9.88{\pm}0.30sec$ for ${\lambda}$ DNA, micrococcus DNA, and T4 DNA, respectively. T4 DNA is trapped in the micropillar array when the crossed electric field of 5V/0.8cm is applied alternately at a 10 second time interval. The present DNA microextractor filters the DNA in a specific size range by adjusting the magnitude and/or the period of the crossed electric field applied in the micropillar array.

Practical Use of DNA Polymorphisms in the Avian Immunoglobulin Light Chain Constant Domain for Species-specific PCR (조류의 종 특이 구별을 위한 항체 유전자의 이용)

  • Choi, J.W.;Kang, S.J.;Park, M.S.;Kim, J.-K.;Han, J.Y.
    • Journal of Animal Science and Technology
    • /
    • v.50 no.1
    • /
    • pp.9-18
    • /
    • 2008
  • Species-specific polymorphisms in chicken, pheasant, turkey, and quail were identified by cloning and sequencing of the immunoglobulin constant domain (IgLC). A set of species-specific primers were then designed on the basis of polymorphisms in the IgLC between species, as well as two additional sets of primers for the cytochrome b and tapasin genes, for the purpose of species identification. Together, the primers successfully distinguished specific species from chicken by species-specific PCR. This simple but unambiguous method may be used to screen avian inter-species germline chimeras, which are valuable models for the conservation of endangered species.