• Title/Summary/Keyword: PCR marker

Search Result 788, Processing Time 0.035 seconds

Refinements for the amplification and sequencing of red algal DNA barcode and RedToL phylogenetic markers: a summary of current primers, profiles and strategies

  • Saunders, Gary W.;Moore, Tanya E.
    • ALGAE
    • /
    • v.28 no.1
    • /
    • pp.31-43
    • /
    • 2013
  • This review provides a comprehensive summary of the PCR primers and profiles currently in use in our laboratory for red algal DNA barcoding and phylogenetic research. While work focuses on florideophyte taxa, many of the markers have been applied successfully to the Bangiales, as well as other lineages previously assigned to the Bangiophyceae sensu lato. All of the primers currently in use with their respective amplification profiles and strategies are provided, which can include full fragment, overlapping fragments and what might best be called "informed overlapping fragments", i.e., a fragment for a marker is amplified and sequenced for a taxon and those sequence data are then used to identify the best primers to amplify the remaining fragment(s) for that marker. We extend this strategy for the more variable markers with sequence from the external PCR primers used to "inform" the selection of internal sequencing primers. This summary will hopefully serve as a useful resource to systematists in the red algal community.

SCAR markers were developed to identify zoysiagrass mutants exhibiting fine leaf characteristics (세엽 한국들잔디 변이체 식별을 위한 SCAR 마커 개발)

  • Chung, Sung Jin;Park, Su Jeong;Choi, Young In;Kim, In-Kyung;Lee, Ka-Yeon;Kim, Hun-Joong;Lee, Geung-Joo
    • Korean Journal of Agricultural Science
    • /
    • v.40 no.2
    • /
    • pp.115-121
    • /
    • 2013
  • Polymorphic bands of two fine-leaf zoysiagrass mutants (CNU 70-1, CNU 70-2) induced via a gamma-ray irradiation on seeds of Zoysia japonica were obtained by using randomly amplified polymorphic DNA (RAPD) primers. The genotype-specific fragments were then converted into PCR-based sequence characterized amplified region (SCAR) markers, which are now amenable to detecting them among other zoysiagrass species widely noticeable in Korea. The CNU 70-1-specific primer set amplified about 900 bp successfully, while the CNU 70-6 marker produced the expected 1,500 bp band, by which those markers were nominated by CNU 70-1_900 and CNU 70-6_1500 SCARs, respectively. The developed SCAR markers can be an applicable tool in sod industry where illegal appropriation hampers breeder's right and profits due to the turfgrass plant vegetatively propagating.

A Case of Parentage Testing in Dog by Microsatellite DNA Typing (Microsatellite DNA형에 의한 개의 친자감정예)

  • 조길재;조병욱;이길왕;김선구;김용균
    • Journal of Life Science
    • /
    • v.13 no.4
    • /
    • pp.416-420
    • /
    • 2003
  • We performed this study to determine the parentage verification of putative dogs. A total of 7 samples (4 Labrador Retriever dog and 3 Poongsan dog) were genotyped by using 12 international markers (PEZ1, PEZ5, PEZ8, PEZ10, PEZ11, PEZ12, PEZ17, PEZ20, PEZ21, FHC2010, FHC2054, FHC2079). This methods consisted of multiplexing PCR procedures, and it showed reasonable amplification of all PCR products. Genotyping was performed with an ABI 310 genetic analyzer. Labrador Retriever Pup I and Pup II were included according to principles of Mendelian genetics in all loci, while Poongsan Pup III was excluded with markers PEZ1 (106/118), PEZ10 (276/300), and FHC2010 (228/232). These results suggest that the present DNA typing is so useful for parentage verification of these two breeds.

Simvastatin Induces Avian Muscle Protein Degradation through Muscle Atrophy Signaling (Simvastatin이 메추리 근육 세포에 미치는 영향)

  • JeongWoong, Park;Yu-Seung, Choi;Sarang, Choi;Sang In, Lee;Sangsu, Shin
    • Korean Journal of Poultry Science
    • /
    • v.49 no.4
    • /
    • pp.265-272
    • /
    • 2022
  • Many studies on poultry have been conducted in the poultry industry to improve their important economic traits, such as egg production, meat quality, and carcass yield. Environmental changes affect the poultry's economic traits, including muscle growth. The purpose of this study is to investigate the mechanisms by which simvastatin causes muscle injury in quail muscle cells. Following treatment with various doses of simvastatin, LD50 in the quail myoblast cells was determined using a cell viability test; cell death was caused by apoptosis and/or necrosis. Thereafter, the expression patterns of the atrophy marker genes were examined via quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The results showed that the transcriptional levels of the muscle atrophy marker genes (Atrogin-1, TRIM63) and the upstream genes in their signaling cascade were increased by simvastatin treatment. This indicated that simvastatin induced myogenic cell death and muscle injury via protein degradation through muscle atrophy signaling. Further studies should focus on identifying the mechanism by which simvastatin induces the protein degradation signaling pathway in quail muscle..

A Novel Marker for the Species-Specific Detection and Quantitation of Shigella sonnei by Targeting a Methylase Gene

  • Cho, Min Seok;Ahn, Tae-Young;Joh, Kiseong;Kwon, Oh-Sang;Jheong, Won-Hwa;Park, Dong Suk
    • Journal of Microbiology and Biotechnology
    • /
    • v.22 no.8
    • /
    • pp.1113-1117
    • /
    • 2012
  • Shigella sonnei is a causal agent of fever, nausea, stomach cramps, vomiting, and diarrheal disease. The present study describes a quantitative polymerase chain reaction (qPCR) assay for the specific detection of S. sonnei using a primer pair based on the methylase gene for the amplification of a 325 bp DNA fragment. The qPCR primer set for the accurate diagnosis of Shigella sonnei was developed from publically available genome sequences. This quantitative PCR-based method will potentially simplify and facilitate the diagnosis of this pathogen and guide disease management.

Quantitation of Hepatitis C Viral RNA Using Direct CRT-PCR

  • Park, Young-Suk;Lee, Kyung-Ok;Oh, Moon-Ju;Chai, Young-Gyu
    • BMB Reports
    • /
    • v.30 no.3
    • /
    • pp.234-236
    • /
    • 1997
  • Chronic hepatitis C virus (HCV) infection is associated with the rapid development of cirrhosis and hepatocellular carcinoma. It has been reported that the amount of HCV RNA may be correlated with the progression of hepatitis and may be a prognostic marker for treatment of HCV patients. The direct detection of HCV RNA by reverse transcription-polymerase chain reaction (RT-PCR) is widely used to determine the presence of circulating virions. The most relevant limit of this approach is the lack of quantitative information about the viral titer. In the present study, we developed the method for HCV quantitation using competitive reverse transcription (CRT)-PCR using the deleted HCV standard. The serially diluted standard was added in titrated amounts to the target HCV RNA. The mixture was then reverse transcribed and amplified in the same reaction tube. The methods were evaluated using over 110 HCV-PCR positive samples in Koreans. About 59% of the samples were judged to contain $10^{5}-10^{6}$ copies of HCV RNA in 1 ml of serum.

  • PDF

Expression of Green Fluorescent Protein(GFP) Gene in Rabbit Embryos (토끼 수정란에서 Green Fluorescent Protein 유전자의 발현)

  • Kang, T. Y.;Yin, X. J.;Chae, Y. J.;Lee, H.;Lee, H. J
    • Journal of Embryo Transfer
    • /
    • v.14 no.1
    • /
    • pp.1-8
    • /
    • 1999
  • The efficiency of transgenic livestock animal production may be improved by early selection of transgenci preimplantation embryos. To examine the possibility of GFP gene as a non-invasive marker for the early screening of transgenic embryo, the GFP gene was microinjected into rabbit zygotes and the later stages of preimplantation embryos were examined for the expression of GFP. The presence of injected DNA was detected by PCR analysis and the expression of GFP was detected by observing green fluorescence in embryos under a fluorescent microscope. Out of 108 GFP gene-injected rabbit zygotes, seventy three(67.6%) were fluorescence-positive. When 11 fluroresecence-positive blastocysts were analyzed for the presence of GFP gene by PCR, 6(54.5%) were positive, and all of the 8 flrouescence-negative blastocysts were also negative by PCR. The results indicate that the screening of transgene in rabbit embryos by PCR analysis and GFP detection could be a promising method for the preselection of transgenic embryos.

  • PDF

PCR-based Assay for the Specific Detection of Pseudomonas syringae pv. tagetis using an AFLP-derived Marker

  • Song, Eun-Sung;Kim, Song-Yi;Chae, Soo-Cheon;Kim, Jeong-Gu;Cho, Heejung;Kim, Seunghwan;Lee, Byoung-Moo
    • Research in Plant Disease
    • /
    • v.21 no.1
    • /
    • pp.1-5
    • /
    • 2015
  • A PCR method has been developed for the pathovar-specific detection of Pseudomonas syringae pv. tagetis, which is the causal agent of bacterial leaf spots and apical chlorosis of several species within the Compositae family. One primer set, PSTF and PSTR, was designed using a genomic locus derived from an amplified fragment length polymorphism (AFLP) fragment produced a 554-bp amplicon from 4 isolates of P. syringae pv. tagetis. In DNA dot-blot analysis with the PCR product as probe, a positive signal was identified in only 4 isolates of P. syringae pv. tagetis. These results suggest that this PCR-based assay will be a useful method for the detection and identification of P. syringae pv. tagetis.

DNA Analysis of mtDNA COI Gene in the Sharp-toothed Eel (Muraenesox cinereus Forskal) from Yeosu, Jinhae, Jeju, Goseoung, Jangheung and Haenam Populations in Korea Using PCR-aided RFLP

  • Oh, Taeg-Yun;Jeong, Sun-Beom;Cho, Eun-Seob
    • Journal of Environmental Science International
    • /
    • v.20 no.4
    • /
    • pp.551-554
    • /
    • 2011
  • The production of the sharp-toothed eel by commercial catch off waters of Korea is annually declined after 1978. This study was carried out to obtain the stock management of the sharp-toothed eel using the PCR-aided RFLP method. The mtDNA COI gene was amplified using species-specific primers and PCR product was observed to 700 bp. Amplified DNA fragments were treated with six kinds of restriction enzymes (BaeHI, EcoRI, PstI, Ksp22, HinfI and HaeIII). The treatment of HaeIII showed a distinct PCR product between Yeosu/Jinhae/Jeju/Goseoung and Jangheung/Haenam populations that were observed from 300 to 400 bp in reference to 100 bp molecular marker. However, DNA fragment within populations had an identical pattern. The phylogenetic homology is 82% between two populations inferred from RFLP PCR product pattern using NTsysPC ver. 2.1. The use of HaeIII plays an important role in discriminating populations. It is thought that adults after over-wintering in the southern part of Jeju migrate to the Yeosu, Jinhae and Goseoung regions to spawn instead of to southwestern waters. Individuals within populations showed a relatively active genetic mixing and migration regardless of geography. However, the genetic ancestor of Jangheung and Haenam populations is appeared to be more adjacent to China or Japan than Jeju.

Species Identification of Five Penaeid Shrimps Using PCR-RFLP and SSCP Analyses of 16S Ribosomal DNA

  • Khamnamtong, Bavornlak;Klinbunga, Sirawut;Menasveta, Piamsak
    • BMB Reports
    • /
    • v.38 no.4
    • /
    • pp.491-499
    • /
    • 2005
  • DNA-based molecular markers for differentiation of five penaeid shrimps (Penaeus monodon, P. semisulcatus, Feneropenaeus merguiensis, Litopenaeus vannamei and Marsupenaeus japonicus) were developed based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and single-stranded conformation polymorphism (SSCP) of 16S ribosomal (r) DNA. Differentiation of P. monodon, P. semisulcatus and L. vannamei can be unambiguously carried out by PCR-RFLP of 16S $rDNA_{560}$ whereas P. semisulcatus and M. japonicus shared a BABB mitotype. These shrimps were successfully discriminated by SSCP analysis of 16S $rDNA_{560}$. Nevertheless, the amplification success for L. vannamei and F. merguiensis was not consistent when tested against larger sample sizes. As a result, 16S $rDNA_{560}$ of an individual representing the most common mitotype of each species was cloned and sequenced. The new primer pair was designed and tested against the large sample sizes (312 bp product, N = 185). The amplification success was consistent across all species. PCR-RFLP of 16S $rDNA_{312}$ was as effective as that of 16S $rDNA_{560}$. Differentiation of all shrimp species were successfully carried out by SSCP analysis.