• 기술혁신학회지
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• 제2권3호
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• pp.94-106
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• 1999

In Korea Basic Science Institute(KBSI) the remote data analysis system is developed for the joint use of advanced equipments. This system enables the researchers to access the datas which are produced at KBSI and analyse them by Java program on the Web,. Except Web browser such as Internet Explorer or Netscape Navigator no additional softwares are required for analysing data. We have developed remote data analysis systems for five major equipments which KBSI supports for the researchers, The systems which are developed are those for NMR spectrometer High Reso-lution Tandem mass Spectrometer Microscopic Imaging System DNA Sequencer and Natural Ra-dioactivity Measruement System, These programs work on any computer platform and any operat-ing system only if the internet is available. This remote data analysis system will be served as a part of Collaboratory the remote collaborative system.

• 농업생명과학연구
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• 제43권2호
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• pp.31-38
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• 2009

DNA sequencer는 template로 이용하는 DNA의 quality와 sequencing 반응 산물의 정제 방법, 그리고 gel 농도에 민감하다고 알려져 있다. 이에 우리는 plasmid DNA의 준비, 정제, sequencing 반응, gel 농도와 injection medium 등에 대한 최적 조건을 구축하기 위한 연구를 수행하였다. Plasmid DNA 준비과정에서 phenol을 사용한 것 보다 chloroform을 사용한 것이 평균 reading length가 532 bp에서 684 bp로 향상되었으며, 2.5% DMSO를 첨가한 것이 첨가하지 않은것에 비해 200 bp 더 길게 염기서열 분석이 되었다. 또한, sequencing 반응산물 정제 시 50 mM EDTA와 0.6 M sodium acetate를 미리 섞어서 pH 8.0으로 맞춘 것을 사용한 것이 50 mM EDTA(pH 8.0)와 0.6 M sodium acetate(pH 5.2)를 각각 사용한 것 보다 20 bp 길게 염기서열 분석이 되었다. Injection medium으로는 실험실에서 resin으로 탈 이온화 시킨 formamide보다 정제된 ABI formamide를 사용한 것이 보다 재현성 있게 reading length가 90 bp 더 길게 분석 되었으며, 4% PAGE gel 보다 3.6% PAGE gel을 사용한 것이 150 bp 더 길게 분석 되었다. Template 준비 시 chloroform으로 정제하고 2.5% DMSO를 첨가, sequencing 반응산물 정제 시 carrier의 pH를 8.0으로 맞춘 것을 이용, 그리고 ABI formamide와 3.6% gel 농도를 사용하는 최적의 조건으로 평균 700 bp, 85% score를 얻을 수 있었다.

• Fisheries and Aquatic Sciences
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• 제9권2호
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• pp.70-74
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• 2006

DNA sequence-based typing is considered a robust tool for the discrimination of dinoflagellate species because of the availability of extensive rDNA sequences. Here, we present a rapid, cost-effective DNA-sequencing technique for various PCR products. This sequencing strategy relies on 'nested' or 'tailed' primer labeled with near-infrared dye, and uses a minimal volume of unpurified PCR product (ca. $5{\mu}L$) as the DNA template for sequencing reactions. Reliable and accurate base identification was obtained for several hundred PCR fragments of rRNA genes. This quick, inexpensive technique is widely applicable to sequence-based typing in clinical applications, as well as to large-scale DNA sequencing of the same genomic regions from related species for studies of molecular evolution.

• 생명과학회지
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• 제2권2호
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• pp.108-119
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• 1992

근년에 개발된 효소적인 DNA증폭법을 이용하면 일차구조상의 단편적인 정보만 알면 단 수시간내 해석에 필요한 양의 DNA가 증폭되어 cDNA의 염기배열결정의 신속화, 간편화가 가능하게 되었다. 그러므로 유전자증폭법으로써 PCR법에 관해 기술한다. 그리고 리보좀 RNA는 분자시계로서 생물의 계통을 논하는 데에 있어서는 최적의 조건을 갖춘 고분자화합물이다. 이에 PCR법을 이용한 16S like 리보좀DNA의 증폭법을 다루고, PCR증폭산물의 염기배열결정법에 대해 서술한다. 또한 인위적인 leading error 등을 배제하고 신속한 자동해독과 시간적인 절약이 자동 DNA sequencer의 개발과 시판으로 가능하게 되어 cDNA의 형광색소표식 염기배열결정법에 대해서도 서술한다.

• 보존과학연구
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• 통권24호
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• pp.153-167
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• 2003

We performed nuclear DNA typing and mitochondrial DNA sequencing analysis based on PCR from an ancient Korean remainsexcavated from Siheung in Korea. 7 bones were collected and partially STR(short tandem repeat) systems, Sex determination Amelogenin kit(Promega co, USA), were used in this study. Mitochondrial DNAs were also amplified and sequenced by ABI 310 DNA sequencer. We know that sample no. 2 and no. 3 were females and also sample no. 2 and no.7 possessed the same maternal inheritance by mitochondrial DNA sequencing results. Throughout this research, the mitochondrial DNA sequencing of human in the middle of Joseon Dynasty in Korea is obtained. In addition, this finding will be an important foundation for the future research.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.25-25
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• 2018

The Fukushima Daiichi Nuclear Power Plant accident in March 2011 caused severe radioactive contamination in the surrounding environment. Since the accident, much attention has been paid to the biological and genetic consequences of organism inhabiting the contaminated area. The effect of radiation exposure on genetic mutation rates is little known, especially for low doses and in situ conditions. Evaluating DNA mutation by low levels of radiation dose is difficult due to the rare mutation event and lack of sequence information before the accident. In this study, correlations with air dose levels and somatic DNA mutation rates were evaluated using Next Generation Sequencer for the clonal plant, Phyllostachys edulis. This bamboo is known to spread an identical clone throughout Japan, and it has the advantage that we can compare genetic mutation rate among identical clone growing different air dose levels. We collected 94 samples of P. edulis from 14 sites with air dose rates from $0.04{\sim}7.80{\mu}Gy/h$. Their clonal identity was confirmed by analysis using 24 microsatellite markers, and then, sequences among samples were compared by MIG sequence. The sequence data were obtained from 2,718 loci. About ~200,000 bp sequence (80 bp X 2,718 loci) were obtained for each sample, and this corresponds to about 0.01% of the genome sequence of P. edulis. In these sequences, 442 loci showed polymorphism patterns including recent origin mutation, old mutation, and sequence errors. The number of mutations per sample ranged from 0 to 13, and did not correlate with air dose levels. This result indicated that DNA mutations have not accumulated in P. edulis living in the air doses levels less than $10{\mu}Gy/h$. Our study also suggests that mutation rates can be assessed by selecting an appropriate experimental approach and analyzing with next generation sequencer.

• 대한한의학방제학회지
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• 제19권2호
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• pp.83-92
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• 2011

Objectives : This study was carried out to moniter microbial flora on freeze-dried herbal powder and identify isolated bacteria. Methods : We measured the total number of bacteria and fungi in 29 herbal powder which had made according to the guideline of KFDA. For the identification, we observed microscopic properties and carried out polymerase chain reaction(PCR). The purified DNA was analyzed by DNA sequencer. Results : Among the 29 herbal powders, the fungi were detected only one sample as unacceptable range of total aerobic bacteria. Isolated bacteria were identified as Bacillus cereus, B. subtilis, B. megaterium, B. licheniformis, Erwinia tasmaniensis, E. amylovora, and Pantoea agglomerans by 16S rDNA analysis. E. tasmaniensis was observed 20 herbal samples. Conclusions : According to above results, further studies for the effective sterilization of low herbal materials should be needed.

• 대한한의학방제학회지
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• 제15권2호
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• pp.119-126
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• 2007

Objectives: This study presents observation of microorganism such as total aerobic bacteria, total fungus, E. coli, Pseudonomas aerugjnosa, Staphylococcus aureus, and Salmonella typhimurium in herbal decoction manufactured by Korean medical clinic. Methods: We examined to observe microorganism using the requirements for the experimental methods recommended by FDA. For the identification, we observed microscopic methods and carried out polymerase chain reaction (PCR) and DNA purification. The purified DNA samples were analyzed by DNA sequencer. As compared with NCBI database. the results were identified by sequences similarity. Results and conclusion: 26 (55%) of 46 decoctions observed positive for microbial test. 12 (46 %) of 26 positive decoctions exceed requirement of microbial limit test. These microbial colony identified genus of Bacillus using microscopic and DNA sequencing methods.

• 보존과학연구
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• 통권23호
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• pp.59-75
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• 2002

In this study human bones and teeth, excavated from the Myeongam-risite in Asan, Chungcheongnam-do Province, have been analysed by nuclear DNA typing and mitochondrial DNA sequencing methods. Twenty-one samples of long bones and twenty-seven samples of teeth from twenty-one individuals were collected and analysed. Among these thirteenteeth were successfully subjected to nuclear DNA extraction, quantification, and PCR(Polymerase Chain Reaction) amplification. Silver STR III (D16S539, D7S820, D13S317) multiplex PCR method was used in this study for a short tandem repeat (STR) analysis. Mitochondrial DNAs of tooth samples were also amplified and sequenced by a DNA sequencer. These analyses show that a sample from Burial no. 29 and one from Burial no. 38(right) possessed the same maternal inheritance. This may suggest that the Myeongam-ri cemetery was used by a kin group for a relatively long period of time.

• Clinical and Experimental Pediatrics
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• 제45권10호
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• pp.1263-1272
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• 2002

목 적 : 레트증후군이란 1966년 Andreas Rett에 의해 처음 기술되었으며 생후 6개월에서 18개월 정도까지 비교적 정상 발달을 한 후 두위 발달의 감소와 함께 습득했던 인지 및 운동능력의 상실, 언어기능의 상실, 그리고 특징적인 손동작(상동행동)을 보이는 X 염색체 우성유전으로 생각되는 질환이다. 1999년 미국에서 그 결함 유전자인 MECP2 유전자가 Xq28에서 밝혀졌으나 우리 나라에서는 이 질환에 대한 전반적 이해가 부족하며 유전자 연구는 거의 전무한 상태이다. 이에 저자들은 우리 나라의 Rett 증후군 환아에서 MECP2 유전자의 결함이 어느 정도 나타나는지를 알고자 이 연구를 시행하였다. 방 법 : 2000년 2월부터 2001년 3월까지 본원을 방문한 Rett 증후군 환아 34례의 말초혈액을 EDTA (Ethylenediaminetetraacetic acid) 처리하여 5 cc 채취하여 냉동 보관한 후 해빙하여 DNA 표본을 추출했다. DNA의 추출은 E.Z.N.A blood DNA kit을 사용하였다. MECP2 유전자 네 종류의 axon은 PCR을 이용해 증폭하였고 primer sequence는 1999년 Amir 등에 의해 디자인 된 것(AF030876)을 사용하였다. ABI 377 DNA sequencer와 ABI PRISM dye cycle sequencing reaction kit을 사용하여 DNA sequencing을 시행하였고 우리 나라에서 최초로 발견된 유전자 변이에 대해서는 RFLP 분석을 통해 확인하였다. 결 과 : 1) MECP2 유전자의 변이를 보인 환아는 23례로 67.6%에서 관찰되었다. 2) 9종의 missense mutation과 3종의 nonsense mutation을 합해 총 12종의 유전자 변이가 관찰되었다. 3) 이들 돌연 변이 중 L100V, G161E, 그리고 T311M은 전세계적으로 우리 나라에서 처음 발견된 변이이다. 4) 23례의 유전자 변이는 대부분(78.3%) MBD와 TRD라는 기능영역에서 발견되었다. 5) 본 연구에서 T158M, R270X, 그리고 R306C가 자주 나타나는 유전자 변이였다. 결 론 : 우리 나라의 Rett 증후군 환아에서도 MECP2 유전자의 변이는 비교적 흔히 관찰되어 MECP2 유전자의 이상이 Rett 증후군을 유발하는 주 유전자 이상임을 확인하였고 Rett 증후군 환아의 확진을 위해 MECP2 유전자 검사가 필요할 것으로 사료된다.