• Title/Summary/Keyword: DNA sequencer

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Genetic Variations of D16S539 Locus in the Korean Population (한국인에서 D16S539 유전좌의 유전적 다형성)

  • Shin, Kyoung-Jin;Yang, Yun-Seok;Choi, Jong-Hoon;Yang, Woo-Ick;Cho, Sang-Ho;Kim, Chong-Youl
    • Journal of Oral Medicine and Pain
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    • v.25 no.4
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    • pp.395-402
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    • 2000
  • The D165539 locus was investigated to collect population genetic data in the Korean population. The selected subject was unrelated 293 Korean people. DNA was extracted from the samples and PCR was performed with fluorescent primer. The amplified fragment was analysed by automated DNA sequencer and it's application software. Among the Korean population, 7 allele and 18 geneotype were observed and allele No. 9 is mostly frequent(0.2679) and then allele No. 11(0.2679), allele No. 9(0.2645). The observed heterozygosity and the expected heterozygosity is 0.7466, 0.7829 each. The polymorphism information content(PIC) is 0.7466. The power of discrimination(PD) and the mean exclusion chance(MEC) are calculated to be 0.9190 and 0.5775.

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Beagle dogs parentage testing by using 22 ISAG microsatellite markers

  • Ji, Hye-jung;Kim, Eun-hee;Lee, Kyoung-kap;Kang, Tae-young;Lee, Joo-myoung;Shin, Hyoung-doo;Kim, Lyoung-hyo;Yun, Young-min
    • Korean Journal of Veterinary Research
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    • v.47 no.4
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    • pp.457-460
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    • 2007
  • The objective of the study was to establish routine parentage testing system in Beagle dogs using 22 ISAG (International Society for Animal Genetics) canine microsatellite markers (2005). Blood collections were obtained from a mother dog, 4 candidate father dogs and 3 offspring (n = 8). Genomic DNA samples were extracted from 8 Beagle dogs blood for PCR analysis. PCR products for the allele were analyzed by ABI 3130 DNA Sequencer and GeneScan (Ver 3.0) analysis and Genotyper (Ver. 2.1) software. The genetic relationship of mother and 3 offspring as well as one father dog among 4 candidate father dogs was confirmed by microsatellite allele analysis. The results of locus for amelogenin, which was designed for sexing, were matching with real gender among 8 Beagle dogs (female; 217/217 homozygosity, male; 179/217 heterozygosity). Twenty two ISAG microsatellite markers are useful the parentage test of Beagle dogs. In addition, amelogenin is an applicable marker to detecting real sex in dogs.

Differential Display Detection of Acid-inducible Genes from Porphyra yezoensis Thalli (해조류 방사무늬김 (Porphra yezoensis) 엽체로부터 산 유도 유전자의 분리)

  • JIN Long-Guo;KANG Se-Eun;CHOI Jae-Suk;PARK Sun-Mee;PARK Jung-Youn;JIN Duck-Hee;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.37 no.4
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    • pp.269-274
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    • 2004
  • Genetic responses of the edible seaweed Porphyra yezoensis tissue to acid shock have been compared using differential display technique. The tissue was challenged in seawater containing $0.05{\%}$ hydrogen chloride (pH 3.0) for 5 min, then rehabilitated in normal seawater for 10 min, 30 min, 60 min and 4 hrs. Total RNA extracted by the LiCl-guanidium method was reverse transcribed and amplified by PCR with arbitrary primers. The amplified fragment responded by the acid shock was selectively isolated from agarose gel and sequenced with DNA auto sequencer. Sequence (1056 bp) of the cDNA contained at least two genes for ASP7K (MW 7418) and ASP5K (MW 5512) proteins.

Analysis of p53 Somatic Mutation in Head and Neck Cancer Using Denaturing High Performance Liquid Chromatography(DHPLC) (두경부 종양에서 DHPLC를 이용한 p53체세포 돌연변이 검출 연구)

  • Kim, Kwang-Youl;Park, Sang-Bum;Han, Sang-Man;Nam, Youn-Hyoung;Jang, Won-Cheoul
    • Journal of the Korean Chemical Society
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    • v.48 no.1
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    • pp.33-38
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    • 2004
  • Mutation of p53 tumor suppressor gene in HNSCC (head and neck squamous cell carcinoma) has been proposed high rate. We extracted genomic DNA from 50 head and neck cancer. The DNA was amplified by PCR at exon 5-8 in p53 tumor suppressor gene. We have compared single strand conformation polymorphism (SSCP) and denaturing high performance liquid chromatography (DHPLC) method for analysis of p53 somatic mutation. As a result, 16 deleted mutations (32%) were detected by SSCP analysis and 17 deleted mutations (34%) were detected by DHPLC analysis at exon 8. All of 17 mutations were proved by sequencing. We conclude that DHPLC is a fast and simple screening method rather than SSCP analysis.

Isolation of an Acid-Labile Gene from the Seaweed Porphyra yezoensis Tissue (해조류 김 Porphyra yezoensis 엽체로부터 산에 민감한 유전자의 분리)

  • Jin, Hyung-Joo;Park, Sun-Mee;Kim, Long-Guo;Jin, Deuk-Hee;Kong, In-Soo;Hong, Yong-Ki
    • KSBB Journal
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    • v.14 no.6
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    • pp.702-706
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    • 1999
  • The genetic responses of aquaculturable seaweed Prophyra yezoensis tissue by acid shock have been compared using differential display technique. The tissue has been challenged in seawater containing 0.05% hydrogen chloride(pH 3.0) for 5 min, then rehabilitated in normal seawater for 10 min, 30 min, 60 min and 4 hrs, respectively. Total RNA was extracted by LiCl-guanidinium method. The cDNA was synthesized by reverse transcription with random hexamers and amplified by PCR with arbitrary primers. The genetic fragment disappeared by acid shock was selectively isolated from agarose gel and sequenced with a DNA auto sequencer. One of the acid-labile gene(605 bp) was identified as a dethiobiotin synthetase gene according to sequence alignment analysis by the NCBI BLAST search program.

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Survey of the Applications of NGS to Whole-Genome Sequencing and Expression Profiling

  • Lim, Jong-Sung;Choi, Beom-Soon;Lee, Jeong-Soo;Shin, Chan-Seok;Yang, Tae-Jin;Rhee, Jae-Sung;Lee, Jae-Seong;Choi, Ik-Young
    • Genomics & Informatics
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    • v.10 no.1
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    • pp.1-8
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    • 2012
  • Recently, the technologies of DNA sequence variation and gene expression profiling have been used widely as approaches in the expertise of genome biology and genetics. The application to genome study has been particularly developed with the introduction of the nextgeneration DNA sequencer (NGS) Roche/454 and Illumina/ Solexa systems, along with bioinformation analysis technologies of whole-genome $de$ $novo$ assembly, expression profiling, DNA variation discovery, and genotyping. Both massive whole-genome shotgun paired-end sequencing and mate paired-end sequencing data are important steps for constructing $de$ $novo$ assembly of novel genome sequencing data. It is necessary to have DNA sequence information from a multiplatform NGS with at least $2{\times}$ and $30{\times}$ depth sequence of genome coverage using Roche/454 and Illumina/Solexa, respectively, for effective an way of de novo assembly. Massive shortlength reading data from the Illumina/Solexa system is enough to discover DNA variation, resulting in reducing the cost of DNA sequencing. Whole-genome expression profile data are useful to approach genome system biology with quantification of expressed RNAs from a wholegenome transcriptome, depending on the tissue samples. The hybrid mRNA sequences from Rohce/454 and Illumina/Solexa are more powerful to find novel genes through $de$ $novo$ assembly in any whole-genome sequenced species. The $20{\times}$ and $50{\times}$ coverage of the estimated transcriptome sequences using Roche/454 and Illumina/Solexa, respectively, is effective to create novel expressed reference sequences. However, only an average $30{\times}$ coverage of a transcriptome with short read sequences of Illumina/Solexa is enough to check expression quantification, compared to the reference expressed sequence tag sequence.

A STUDY ON MUTATIONS OF P53 TUMOR SUPPRESSOR GENE IN ORAL TUMORS (구강종양에서 p53 종양억제 유전자의 돌연변이에 관한 연구)

  • Joo, Seong-Chai;Pyo, Sung-Woon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.1
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    • pp.45-52
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    • 2000
  • Nowadays, there are a lot of evidence that mutation of the p53 tumor suppressor gene is one of the most common genetic abnormalities in neoplastic progression. In this study, we analyzed 20 specimens of oral tumors(squamous cell carcinoma 14 cases, ameloblastoma 3 cases, adenoid cystic carcinoma 2 cases, malignant schwannoma 1 case)using polymerase chain reaction and direct sequencing which used an automated DNA sequencer and software for detection of mutations. Polymerase chain reactions were performed with 4 sets of primers encompassing exon 5, 6, 7, 8, and direct sequencing method was employed. The results were as followings. 1. We detected 10 point mutations out of 20 specimens (50%). 2. The genetic alterations included 7 mis-sense mutations resulting in single amino acid subtitutions, 2 silent mutations, 1 non-sense mutations encoding a stop codon. 3. Mutations were mostly in exon 7(7 out of 10 mutations, 70%) and involved codons 225, 234, 235, 236, 238, 247. 4. Therse were 4 cases of $T{\rightarrow}A$ transversion, 2 cases of $C{\rightarrow}A$ transversion, $A{\rightarrow}G$ transition, 1 case of $C{\rightarrow}G$, $T{\rightarrow}G$ transversion respectively. 5. We could find out point mutations more conveniently using PCR - Automated Direct Sequencing method.

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SNPchaser : A Web-based Program for Detecting SNPs Substitution and Heterozygosity Existence (SNPchaser : DNA서열의 SNPs 치환 및 Heterozygosity 확인 프로그램)

  • Jang, Jin-Woo;Lee, Hyun-Chul;Lee, Myung-Hoon;Choi, Yeon-Shik;Choo, Dong-Won;Park, Kie-Jung;Lee, Dae-Sang
    • KSBB Journal
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    • v.24 no.4
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    • pp.410-414
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    • 2009
  • Single-nucleotide polymorphisms (SNPs) are the DNA sequences difference among the same species in the level of nucleic acids and are widely applied in clinical fields such as personalized medicine. The routine and labor-intensive methods to determine SNPs are performing the sequence homology search by using BLAST and navigating the trace of chromatogram files generated by high-throughput DNA sequencing machine by using Chromas program. In this paper, we developed SNPchaser, a web-based program for detecting SNPs substitution and heterozygosity existence, to improve the labor-intensive method in determining SNPs. SNPchaser performed sequence alignment and visualized the suspected region of SNPs by using user's reference sequence, AB1 files, and positional information of SNPs. It simultaneously provided the results of sequences alignment and chromatogram of relevant area of SNPs to user. In addition, SNPchaser can easily determine existence of heterozygosity in SNPs area. SNPchaser is freely accessible via the web site http://www.bioinformatics.ac.kr/SNPchaser and the source codes are available for academic research purpose.

Detection of Mycoplasmas DNA in the Cancer and the Normal Tissues from the Patients with Gastric and Colon Cancer (위암 및 결장암 조직과 그 주변의 정상조직에서 Mycoplasmas DNA의 정색)

  • Chang, Myung-Woong;Shin, Hyun-Chul;Park, In-Dal;Kim, Kwang-Hyuk
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.279-285
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    • 2007
  • Detection of Mycoplasma DNA from the 30 cases of cancer tissues and the normal tissues surrounding the cancer tissues obtained from the patients with gastric cancer and the other 30 cases of cancer tissues and the normal tissues surrounding the cancer tissues obtained from the patients with colon cancer were evaluated by polymerase chain reaction(PCR). The PCR products were sequenced using an ABI 377 automatic DNA sequencer, and these sequences were confirmed by comparing sequences with the database of the National Center for Biotechnology Information BLAST network server. Mycoplasmas DNA were defected in 18 (60%) cases of normal tissues which were around gastric cancer and were 13 (43.3%) cases of gastric cancer tissues. Mycoplasmas DNA were detected in 15(50%) cases of normal tissues which were around colon cancer and 12 (40%) cases of colon cancer tissues. The M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, and M. conjunctivae were detected from the gastric cancer tissues. The M. faucium, M. subdolum,, M. salivarium, M. auris, M. hyosynoviae, M. bovigenitalium and M. pulmonis were detected from the normal tissues around gastric cancer. The M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, M. synoviae M. bovigenitalium, M. gallinarum, and M. moatsii were detected from the colon cancer. The M. faucium, M. subdolum, M. salivarium, M. auris, M. hyosynoviae, M. bovis, M. opalescens, M. bovigenitalium, M. gallinarum, and M. moatsii were detected from the normal tissues around the colon cancer. These results suggest that Mycoplasmas infection may not correlate with gastric cancer and colon cancer, because of the detection rate of Mycoplasmas DNA were not significantly differences between normal and cancer tissues from the patients.

Occurrence of Plant Parasitic Nematodes in Chrysanthemum and ITS and D3-28S rDNA Characterization of Pratylenchus spp. (국화재배지의 식물기생선충 분포조사 및 뿌리썩이선층의 ITS와 D3-28S rDNA 특성조사)

  • Han, Hye-Rim;Lee, Jae-Kook;Choi, Dong-Ro;Han, Man-Jong;Park, Byeong-Yong
    • Korean journal of applied entomology
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    • v.45 no.3 s.144
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    • pp.293-299
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    • 2006
  • A survey was conducted to find out the major plant parasitic nematode in Chrysanthemum morifolium fields in Korea from May to June in 2005. A genus of Pratylenchus was determined as the most important plant parasitic nematode based on analysis of total 50 samples from 8 cities of chrysanthemum field. Pratylenchus showed 86% occurrence rate and average numbered 1,095 per 200cc soils and 1g root. Five Pratylenchus isolates, 'Muan', 'Masan', 'Tean', 'Gumi', 'Jeongup', were selected for the molecular identification of the species of Pratylenchus, and ITS and D3-28S ribosomal DNA were amplified by PCR. For the ITS, only 'Muan' isolate was differentiated by total 1 kb PCR amplification, which was 200 bp larger than all the other isolates. There was no size variation in amplified D3-28S rDNA and all isolate represented approximately 320 bp of PCR product. Sequence data of D3-28S rDNA were analysed by MegAlign program in DNASTAR software and phylogenetic tree was constructed. Sequence homology was 100% between 'Gumi' isolate and 'Tean' isolate and 'Jeongup' isolate was also close to these isolates by 99.7% sequence homology. 'Gumi', 'Tean' group and 'Jeongup' isolate were determined to be closely related to Pratylenchus vulnus by 96.7% and 96.3% similarity in respectively. D3 sequence of 'Masan' isolate was 100% identical to P. penetrans, and 'Muan' isolate showed 99.7% similarity to P. brachyurus. This result was congruent with the branch divergence pattern shown in phylogenetic tree.