• Title/Summary/Keyword: PCR-RAPD marker

Search Result 86, Processing Time 0.024 seconds

Development of RAPD-Derived SCAR Markers and Multiplex-PCR for Authentication of the Schisandrae Fructus (오미자 (五味子) 종 감별을 위한 RAPD 유래 SCAR Marker 및 Multiplex-PCR 기법 개발)

  • Lee, Young Mi;Moon, Byeong Cheol;Ji, Yunui;Seo, Hyeong Seok;Kim, Ho Kyoung
    • Korean Journal of Medicinal Crop Science
    • /
    • v.21 no.3
    • /
    • pp.165-173
    • /
    • 2013
  • The fruits of Schisandra chinensis have been used as an edible ingredient and traditional medicine in Korea. Due to morphological similarities of dried mature fruits, the correct identification of S. chinensis from other closely related Schisandrae species is very difficult. Therefore, molecular biological tools based on genetic analysis are required to identify authentic Schisandrae Fructus. Random amplifed polymorphic DNA (RAPD) and Sequence Characterized Amplified Region (SCAR) were used to develop an easy, reliable and reproducible method for the authentication of these four species. In this paper, we developed several RAPD-derived species specific SCAR markers and established a multiplex-PCR condition suitable to discriminate each species. These genetic markers will be useful to distinguish and authenticate Schisandrae Fructus and four medicinal plants, S. chinensis, S. sphenanthera, S. repanda and K. japonica, in species level.

Detection of DNA Fragment to Differentiate Korean Cattle

  • Yeo, J.S.;Kim, J.W.;Chang, T.K.;Nam, D.H.;Han, J.Y.;Choi, C.B.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.15 no.8
    • /
    • pp.1071-1075
    • /
    • 2002
  • In order to identify and develop the specific DNA marker for the identification of Hanwoo (Korean Cattle) from other breeds, a specific DNA marker of 519 bp was identified and sequenced from polymorphic analysis using RAPD-PCR for 6 cattle breeds. Two different repetitive sequences, $(AAC)_5$ and $(GAAGA)_2$, were selected and designed to use specific probe to develop a DNA marker for Hanwoo specific. When the $(AAC)_5$ probe was applied, the 10 kb specific DNA marker showed in the DNA fingerprinting from 237 of 281 Hanwoo individuals. This novel Hanwoo specific DNA probe is useful to perform the marker-assisted selection for screening Hanwoo purity as an unique genetic source.

Purity Test of Radish Hybrid Seeds Using Randomly amplified Polymorphic DNA Marker

  • Oh, Sei-myoung;Soontae Kwon
    • Journal of Life Science
    • /
    • v.11 no.1
    • /
    • pp.65-67
    • /
    • 2001
  • In order to develop a rapid and simple method for testing the purity of radish hybrid seeds using a procedure based on the PCR(Polymerase chain reaction), eighty random primers were screened with the genomic DNA extracted from five day old seedlings of inbred parent lines and their F1 hybrids. Two primers, HRM-02 (5'-GAGACCAGAC-3') and HRM-19(5'-TGAGGCGTGT-3'), generate reproducible unique PCR patterns which can identify each parent lines as well as their hybrids. In actual test of randomly selected hybrid seeds using the two marker primers, the purity tested by one primer was exactly same as that of other primer. It suggests that one marker primer selected in this experiment is enough for the purity test of radish hybrid seeds. We demonstrates the use of RAPD(randomly amplified polymorphic DNAs) markers to identify each of inbred parent lines and hybrids by rapid and simple method.

  • PDF

Genetic Similarity in Crucian Carp(Carassius carassius) by PCR-RAPD Analysis (PCR-RAPD 분석에 의한 붕어(Carassius carassius)의 유전적 유사성)

  • Yoon, Jong-Man;Kim, Jong-Yeon
    • Development and Reproduction
    • /
    • v.5 no.2
    • /
    • pp.151-158
    • /
    • 2001
  • Genomic DNA from the blood of crucian carp(Carassius carassiu) from lake and aquaculture facility in Kunsan, Korea was extracted in order to identify genetic differences by polymerase chain reaction-randomly amplified polymorphic DNAs(PCR-RAPD). Out of 12 primers, 6 generated 266 highly reproducible RAPD markers, producing approximately 2.1 polymorphic bands per primer in crucian carp from lake. The degree of similarity varied from 0.18 to 0.76 as calculated by bandsharing analysis in crucian carp from lake. The RAPD outlines obtained with DNA of two different crucian carp populations from Kunsan were different(0.47 from lake and 0.70 from aquaculture facility, respectively). The electrophoretic analysis of polymerase chain reaction-randomly amplified polymorphic DNAs(PCR-RAPD) products showed high levels of similarity between different individuals in crucian carp from aquaculture facility. This result implies the genetic similarity due to raising in the same environmental condition or inbreeding within the crucian carp from aquaculture facility in Kunsan. In other words, crucian carp may have high levels of genome DNA diversity due to the introduction of the wild population from the other sites of Knsan even if it may be the geographical diverse distribution of this species. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of important aquacultural fish species, crucian carp. However, in future, additional populations and sampling sites will be necessary to complement weak points.

  • PDF

Molecular Typing of Leuconostoc citreum Strains Isolated from Korean Fermented Foods Using a Random Amplified Polymorphic DNA Marker

  • Kaur, Jasmine;Lee, Sulhee;Sharma, Anshul;Park, Young-Seo
    • Food Engineering Progress
    • /
    • v.21 no.2
    • /
    • pp.174-179
    • /
    • 2017
  • For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Development of Sequence Characterized Amplified Regions (SCAR) Showing for Cheju Native Horse (품종 특이성을 이용한 제주마 판별 표지인자 재발)

  • Cho Byung Wook
    • Journal of Life Science
    • /
    • v.15 no.3 s.70
    • /
    • pp.474-478
    • /
    • 2005
  • This study was conducted to analyze genetic characteristics and to develop the specific marker for Cheju native horse (Coo) at the level of sequence characterized amplified regions (SCARs). We collected blood samples from Cheju native horse and Thoroughbred horse (Th) and obtained genomic DNA from the blood of 50 individuals randomly selected within the breeds. Seven hundred primers were chosen randomly and were used to examin the polymorphism and 40 kinds of primers showed polymorphic RAPD band patterns between two breeds. Thirty primers of them showed horse specific bands. With the primer MG 30, amplified band of 2.0 kb showed the specificity to Cheju native horse (Cnh). Additionally MG 53 detected the thoroughbred horse (Th) specific markers at size of 2.3 kb. As the next, 2.3 kb band from MG 53 was checked with the all individuals from all the breeds of this study, and it maintained the reproducible breed specificity to thoroughbred horse (Th). With this results, 2.3 kb band was cloned into plasmid vector and sequenced bidirectionally from both ends of the cloned fragment. With the obtained sequences 10 nucleotide extended primers including the original arbitray primer were designed as a SCARs primer. Finally, the primer with extended sequence showed the reproducible breed differentiation pattern and it was possible to identify Cheju native horse (Cnh) from other breeds. The SCARs marker 2.3 kb from MG 53 could be used to identify Cheju native horse (Cnh) for not only registration but also horse breeding programe.

Genetic Variations and Relationships of Persicaria thunbergii(Sieb. & Zucc.) H. Gross ex Nakai(Polygonaceae) by the RAPD Analysis (RAPD 분석에 의한 고마리(마디풀과)의 유전적 변이 및 유연관계)

  • Kim, Yong-Hyun;Tae, Kyoung-Hwan;Kim, Joo-Hwan
    • Korean Journal of Plant Resources
    • /
    • v.21 no.1
    • /
    • pp.66-72
    • /
    • 2008
  • RAPD analyses were performed from twenty-four populations of Persicaria thunbergii(Sieb. & Zucc.) H. Gross ex Nakai. The length of amplified DNA fragments ranged from 200 to 1,900bp. 184 scorable RAPD markers were found from PCR reactions with sixteen random oligoprimers. Based on the results, populations of Persicaria thunbergii were classified into disturbance streams of urban and rural streams as well as natural streams. And the populations from natural streams showed having higher genetic similarites than those from highly disturbed streams, Also, the heterogenetic differences between the populations from natural and disturbed areas could be represented the results of the stream environmental changes.

Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains with higher β-glucan (베타글루칸 함량이 높은 큰느타리버섯 선발을 위한 SCAR marker 개발)

  • Kim, Su Cheol;Kim, Hye Soo;Cho, Yong Un;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
    • /
    • v.13 no.1
    • /
    • pp.79-83
    • /
    • 2015
  • In this study, SCAR marker that differentiates Pleurotus eryngii strains with higher ${\beta}$-glucan from control strain was developed. Genomic DNAs of 9 control strains of Pleurotus eryngii and 9 Pleurotus eryngii strains with higher ${\beta}$-glucan were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). One-hundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 91 bp was yielded by OP-R03 primer from the Pleurotus eryngii strains with higher ${\beta}$-glucan. A sequence characterized amplified region (SCAR) marker, designated as OP-R03-1-F and OP-R03-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-R03-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains with higher ${\beta}$-glucan from the control strains.

Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains adaptable to high-temperature (큰느타리버섯의 고온적응성 형질에 관련된 SCAR Marker 개발)

  • Kim, Su Cheol;Kim, Hye Soo;Park, So Yeon;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
    • /
    • v.12 no.3
    • /
    • pp.226-231
    • /
    • 2014
  • In this study, SCAR marker that differentiates Pleurotus eryngii strains adaptable to high-temperature from control strain was developed. Genomic DNAs of 7 control strains of Pleurotus eryngii and 7 Pleurotus eryngii strains adaptable to high-temperature were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). Onehundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 385 bp was yielded by OP-A06 primer from the Pleurotus eryngii strains adaptable to high-temperature. A sequence characterized amplified region (SCAR) marker, designated as OP-A06-1-F and OP-A06-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-A06-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains adaptable to high-temperature from the control strains.

Relationship of Lycoris (Amaryllidaceae) Based on RAPD Markers (RAPD markers에 의한 상사화속 식물의 유연관계)

  • Tae, kyoung-Hwan;Kim, Yong Hyun;Shin, Young-Hwa;Kang, Shin-Ho;Kim, Joo-Hwan;Ko, Sung-Chul
    • Korean Journal of Plant Taxonomy
    • /
    • v.38 no.1
    • /
    • pp.17-29
    • /
    • 2008
  • Phylogenetic relationships were examined for 17 taxa of Lycoris by RAPD analysis. The length of the amplified DNA fragments ranged from 300 bp to 1,700 bp. 57 scorable RAPD markers were observed from PCR reactions with five random oligoprimers. The analysis by UPGMA sepatated the examined taxa of Lycoris into were clusters. First group was comprised of ten taxa of L. chinensis var. sinuolata, L. sanguinea var. koreana, L. uydoensis, L. flavescens, L. radiata var. pumila, L. radiata, L. squamigera, L. chejuensis, L. aurea and L. guangxiensis, second group of L. haywardii, L. sprengeri, L. rosea, L. straminea and L. houdyshii, third group of outgroup of Narcissus tazetta var. chinensis and Crinum asiaticum var. japonicum. From the viewpoint of cytological characters such as polyploidy and karyotype, the RAPD analysis was very useful to show the relationship among the intraspecific taxa of Lycoris.