scholarly journals  Rapid sexing of selected Galliformes by polymerase chain reaction

2012 ◽  
Vol 57 (No. 4) ◽  
pp. 187-192 ◽  
Author(s):  
J. Kalina ◽  
J. Mucksová ◽  
H. Yan ◽  
P. Trefil
Keyword(s):  
Polymerase Chain Reaction ◽  
Meleagris Gallopavo ◽  
Single Pair ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Game Birds ◽  
Commercial Chicken ◽  
Commercial Species ◽  
Common Pheasant ◽  
Polymerase Chain

Vent sexing of one-day-old chicks in commercial hatcheries has long been common practice and can be highly accurate. However, there are circumstances when this technique is not applicable such as smaller breeds, non-domestic birds, or where is the necessity of precise sexing. In this study we present a simple and reliable method for fast gender determination in selected Galliformes for which phenotypic determination of sex is difficult until maturity. Four species were tested: two commercial species – chicken (Gallus gallus) and turkey (Meleagris gallopavo), and two game birds – common pheasant (Phasianus colchicus) and wood grouse (Tetraro urogallus). DNA was tested with universal single-pair primers polymerase chain reaction (PCR) detecting W chromosome specific sequence yielding a single band of length specific for each species. The method was developed with regards to time consumption and cost-effectiveness giving results in less than two hours. The method may also be used for early sexing in commercial chicken and turkey flocks as well as sexing of smaller game birds flocks or for research laboratories when rapid sexing of selected Galliformes cells is required.  

Relationships among blueberry species within the section Cyanococcus of the Vaccinium genus based on EST-PCR markers

2021 ◽  
Author(s):  
Lisa Jeannine Rowland ◽  
Elizabeth L. Ogden ◽  
James R. Ballington
Keyword(s):  
Polymerase Chain Reaction ◽  
Expressed Sequence Tag ◽  
Clustering Methods ◽  
Polyploid Species ◽  
Pcr Markers ◽  
Chain Reaction ◽  
Ploidy Levels ◽  
Commercial Species ◽  
Polymerase Chain ◽  
Expressed Sequence

Commercial blueberry species of North America belong to the Vaccinium genus, section Cyanococcus. Phylogenetic relationships of 50 accessions of different ploidy levels within Cyanococcus were investigated using 249 expressed sequence tag-polymerase chain reaction markers and standard clustering methods. Of the commercial species, tetraploid V. corymbosum grouped most closely with the diploids, V. fuscatum and V. caesariense, followed by the diploid V. elliottii. Tetraploid V. angustifolium grouped with the diploids, V. boreale and V. myrtilloides. Hexaploid V. virgatum grouped most closely with the diploid V. tenellum, thus shedding light on the origins of these polyploid species.

scholarly journals Extração de DNA a partir de sangue humano coagulado para aplicação nas técnicas de genotipagem de antígenos leucocitários humanos e de receptores semelhantes à imunoglobulina

2009 ◽  
Vol 42 (6) ◽  
pp. 651-656 ◽  
Author(s):  
Daniela Maira Cardozo ◽  
Gláucia Andréia Guelsin ◽  
Samaia Laface Clementino ◽  
Fabiano Cavalcante de Melo ◽  
Marco Antônio Braga ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Killer Cell ◽  
Human Leukocyte ◽  
Reaction Sequence ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Salting Out ◽  
Leukocyte Antigens ◽  
Polymerase Chain ◽  
Made In

O objetivo deste estudo foi padronizar uma metodologia de extração de DNA de alta qualidade a partir de amostras de sangue coagulado. Quarenta e oito amostras de sangue humano coagulado foram utilizadas para a extração de DNA pelo kit comercial EZ-DNA® (Biological Industries, Beit Haemek, Israel), pelo kit de coluna Neoscience® (One Lambda Inc., San Diego, CA) e pelo método modificado de salting out. Apenas o método de salting out foi capaz de extrair altas concentrações de DNA (média, 180ng/µL), as quais foram medidas pelo detector de fluorescência Qubit® (Invitrogen, USA). Este método permitiu a amplificação dos genes HLA (human leukocyte antigens) pela tecnologia PCR-SSO (polymerase chain reaction - specific sequence of oligonucleotides) Luminex, a qual exige DNA de boa qualidade, e de genes KIR (killer cell immunoglobulin-like receptors) pela técnica made in house PCR-SSP (polymerase chain reaction-sequence specific of primers), a qual demanda uma concentração específica de DNA (10ng/µL). Concluímos que a técnica de salting out modificada foi muito eficiente, simples e rápida para a extração de DNA de amostras de sangue humano coagulado, com o objetivo de realizar a genotipagem de genes HLA e KIR.

scholarly journals Ligation-Mediated Polymerase Chain Reaction Detection of 8-Oxo-7,8-Dihydro-2′-Deoxyguanosine and 5-Hydroxycytosine at the Codon 176 of the p53 Gene of Hepatitis C-Associated Hepatocellular Carcinoma Patients

2020 ◽  
Vol 21 (18) ◽  
pp. 6753
Author(s):  
Andrea Galli ◽  
Armelle Munnia ◽  
Filippo Cellai ◽  
Mirko Tarocchi ◽  
Elisabetta Ceni ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Polymerase Chain Reaction ◽  
Hepatitis C ◽  
Molecular Mechanisms ◽  
Active Oxygen Species ◽  
Dietary Exposure ◽  
P53 Gene ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Polymerase Chain

Molecular mechanisms underlying Hepatitis C virus (HCV)-associated hepatocellular carcinoma (HCC) pathogenesis are still unclear. Therefore, we analyzed the levels of 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) and other oxidative lesions at codon 176 of the p53 gene, as well as the generation of 3-(2-deoxy-β-d-erythro-pentafuranosyl)pyrimido[1,2-α]purin-10(3H)-one deoxyguanosine (M1dG), in a cohort of HCV-related HCC patients from Italy. Detection of 8-oxodG and 5-hydroxycytosine (5-OHC) was performed by ligation mediated-polymerase chain reaction assay, whereas the levels of M1dG were measured by chromatography and mass-spectrometry. Results indicated a significant 130% excess of 8-oxodG at –TGC– position of p53 codon 176 in HCV-HCC cases as compared to controls, after correction for age and gender, whereas a not significant increment of 5-OHC at –TGC– position was found. Then, regression models showed an 87% significant excess of M1dG in HCV-HCC cases relative to controls. Our study provides evidence that increased adduct binding does not occur randomly on the sequence of the p53 gene but at specific sequence context in HCV-HCC patients. By-products of lipid peroxidation could also yield a role in HCV-HCC development. Results emphasize the importance of active oxygen species in inducing nucleotide lesions at a p53 mutational hotspot in HCV-HCC patients living in geographical areas without dietary exposure to aflatoxin B1.

Comparative analysis of microsatellite loci in chicken and turkey

Genome ◽  
2000 ◽  
Vol 43 (5) ◽  
pp. 796-802 ◽  
Author(s):  
Kent M Reed ◽  
Kristelle M Mendoza ◽  
Craig W Beattie
Keyword(s):  
Polymerase Chain Reaction ◽  
Comparative Analysis ◽  
Microsatellite Markers ◽  
Genomic Dna ◽  
Annealing Temperature ◽  
Meleagris Gallopavo ◽  
Allelic Polymorphism ◽  
Chain Reaction ◽  
Flanking Sequences ◽  
Polymerase Chain

Cross-species amplification of 520 chicken microsatellite markers was tested by polymerase chain reaction with genomic DNA of the turkey (Meleagris gallopavo). Each primer pair was tested at six different combinations of annealing temperature and MgCl2 concentration. A total of 280 (54%) of the primer pairs produced amplification products. The majority of these products were similar, if not identical in size to those expected based on the fragment sizes of the corresponding chicken loci. Structure of the dinucleotide repeat and flanking sequences was examined for 13 turkey fragments (amplified with chicken primers) and 5 chicken fragments (amplified with turkey primers). Sequence analysis found a wide array of mutations between species in addition to differences in repeat length. To estimate the usefulness of the amplified loci for genetic mapping in the turkey, allelic polymorphism was determined for 57 of the 280 amplified loci. A total of 20 of 57 markers (35%) were polymorphic with an average of 1.4 alleles per locus. The results of this study suggest that approximately 20% of the chicken microsatellite markers will be useful for mapping the turkey genome.Key words: microsatellite, chicken, turkey, Meleagris gallopavo.

scholarly journals Evidence of Genetic Diversity Underlying Rh D−, Weak D (Du), and Partial D Phenotypes as Determined by Multiplex Polymerase Chain Reaction Analysis of the RHD Gene

Blood ◽  
1997 ◽  
Vol 89 (7) ◽  
pp. 2568-2577 ◽  
Author(s):  
Neil D. Avent ◽  
Peter G. Martin ◽  
Sylvia S. Armstrong-Fisher ◽  
Wendy Liu ◽  
Kirstin M. Finning ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Basis ◽  
Gene Deletion ◽  
Stop Codon ◽  
Point Mutations ◽  
Polymerase Chain Reaction Analysis ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Multiplex Pcr Assay ◽  
Polymerase Chain

Abstract The human blood group Rh antigens are expressed by proteins encoded by a pair of highly homologous genes located at chromosome 1p34-36. One of the genes (RHCE ) encodes Rh CcEe antigens, while the other (RHD) the D antigen. Point mutations in the RHCE gene generate the C/c and E/e polymorphisms, while it has been shown that an RHD gene deletion can generate the D-negative phenotype. We have analyzed intron 4 of the RHCE and RHD genes and have defined the site of an RHD-specific deletion located in this intron. Using a multiplex RHD typing assay, which combines a reverse polymerase chain reaction (PCR) primer, which straddles this RHD-specific sequence, and a pair of primers located in exon 10 of the RHD gene, we have analyzed 357 different genomic DNA samples derived from individuals expressing D+, D−, weak D, and partial D phenotypes. Of these, we have noted a significant discordance with our multiplex PCR assay in the D− phenotypes dCcee and dccEe (which have been previously described) and weak D phenotypes. Our results suggest that in five serologically D− individuals we have identified an apparently intact RHD gene. Sequence analysis of transcripts obtained from one of these individuals (of phenotype dCCee) illustrates the presence of full-length RHD transcripts, which have a point mutation at nucleotide 121 (C → T), which generates an in-frame stop codon (Gln41Stop). Thus, we describe a different molecular basis for generating the D− phenotype to the complete RHD gene deletion described previously. We also show that there are discordances with serotype and the multiplex assay in weak D and partial D phenotypes, indicating that the underlying molecular basis can be heterogeneous. Existing Rh D PCR assays assume the complete absence of the RHD gene in D− phenotypes. We describe a different molecular basis for generating the D− phenotype to the complete RHD gene deletion described previously.

scholarly journals Novel TaqMan real-time polymerase chain reaction assay for verifying the authenticity of meat and commercial meat products from game birds

2010 ◽  
Vol 27 (6) ◽  
pp. 749-763 ◽  
Author(s):  
María Rojas ◽  
Isabel González ◽  
Miguel Ángel Pavón ◽  
Nicolette Pegels ◽  
Adriana Lago ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Polymerase Chain Reaction Assay ◽  
Meat Products ◽  
Chain Reaction ◽  
Game Birds ◽  
Polymerase Chain

scholarly journals DETECTION OF SALMONELLA SPP. IN COMMERCIAL EGGS FROM LAYER CHICKEN FARMS AND TRADITIONAL MARKETS IN BALI

2021 ◽  
Vol 22 (1) ◽  
pp. 93-100
Author(s):  
Alifianita Anake Yansri ◽  
Hani Plumeriastuti ◽  
Mustofa Helmi Effendi
Keyword(s):  
Polymerase Chain Reaction ◽  
Salmonella Enteritidis ◽  
Chain Reaction ◽  
Salmonella Spp ◽  
Traditional Markets ◽  
Layer Chicken ◽  
Commercial Chicken ◽  
Polymerase Chain ◽  
Duplex Polymerase Chain Reaction ◽  
Chicken Eggs

This study aims to early detect Salmonella spp. and identify serotypes in commercial chicken eggs from layer chicken farms and traditional markets in Bali. Early detection study of Salmonella spp. was carried out by conventional bacteriological methods, while serotype identification by duplex Polymerase Chain Reaction (d-PCR) test against the invA gene from Salmonella spp. and the sefA gene from Salmonella enteritidis. Egg samples in this study were taken from 10 layer chicken farms in Bali which included districts of Bangli, Gianyar, Tabanan and Karangasem. Egg samples from traditional markets were taken from 18 traditional markets from the districts of Bangli, Gianyar, Tabanan, Karangasem, Badung, and Denpasar City. Samples were eggshells and egg whites. Analysis of positive results from Salmonella spp. described descriptively. The results showed that eggshells and white eggs from all of the layer chicken farms are negative contaminated with Salmonella spp. (0%). In eggshell samples taken from the traditional markets of Taman Bali and Tulikup from the districts of Bangli and Gianyar, positive with Salmonella spp. (11,1%) by conventional bacteriological tests. In the duplex Polymerase Chain Reaction test, S. enteritidis serotypes were identified. The finding contamination of Salmonella enteritidis in commercial chicken eggs from traditional markets require periodically detection to prevent the occurrence of salmonellosis due to consumption of contaminated chicken eggs in traditional markets in Bali.

Localization of Tomato Spotted Wilt Virus (Genus Tospovirus, Family Bunyaviridae) in Peanut Pods

1999 ◽  
Vol 26 (2) ◽  
pp. 98-100 ◽  
Author(s):  
S. S. Pappu ◽  
H. R. Pappu ◽  
A. K. Culbreath ◽  
J. W. Todd
Keyword(s):  
Polymerase Chain Reaction ◽  
Immunosorbent Assay ◽  
Tomato Spotted Wilt Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Specific Antibodies ◽  
Tomato Spotted Wilt ◽  
Polymerase Chain ◽  
Wilt Virus

Abstract The localization of tomato spotted wilt virus (TSWV, genus Tospovirus, family Bunyaviridae) in peanut pods was determined by enzyme-linked immunosorbent assay (ELISA) using TSWV specific antibodies. Pods were collected from symptomatic and asymptomatic fieldgrown plants. All the plants were tested by ELISA for presence or absence of TSWV infection. Normal and abnormal looking pods from symptomatic plants were assayed by ELISA. Each pod was divided into shell, testa, and cotyledons. All of the shell and testa samples of both normal and abnormal pods from symptomatic plants were positive for TSWV, whereas TSWV could not be detected in the cotyledons. Similar results were observed by polymerase chain reaction, except that the cotyledons occasionally had a TSWV-specific sequence. No virus could be detected in any part of the pod collected from asymptomatic, virus-free plants. In growout tests of seed from both symptomatic and asymptomatic plants, none of the plants showed TSWV infection when assayed by ELISA. Results demonstrated the preferential accumulation of the virus in the shell and testa.

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