scholarly journals Direct sequencing of a HLA-DRB gene by polymerase chain reaction: Sequence variation in DRw8 specificity

1990 ◽  
Vol 35 (2) ◽  
pp. 151-157 ◽  
Author(s):  
Yoshihisa Watanabe ◽  
Katsushi Tokunaga ◽  
Kazumasa Matsuki ◽  
Keiichi Omoto ◽  
Takeo Juji
Keyword(s):  
Polymerase Chain Reaction ◽  
Sequence Variation ◽  
Direct Sequencing ◽  
Reaction Sequence ◽  
Chain Reaction ◽  
Drb Gene ◽  
Polymerase Chain

A preliminary survey of mitochondrial sequence variation in Triatominae (Hemiptera: Reduviidae) using polymerase chain reaction-based single strand conformational polymorphism (SSCP) analysis and direct sequencing

1998 ◽  
Vol 88 (5) ◽  
pp. 553-560 ◽  
Author(s):  
J. R. Stothard ◽  
Y. Yamamoto ◽  
A. Cherchi ◽  
A. L. Garcia ◽  
S. A. S. Valente ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sequence Variation ◽  
Direct Sequencing ◽  
Single Strand ◽  
Polymorphism Analysis ◽  
Chain Reaction ◽  
Single Strand Conformational Polymorphism ◽  
Conformational Polymorphism ◽  
Polymerase Chain ◽  
Selection Of Species

AbstractGenetic variation within triatomine bugs was investigated by amplification of a 400 bp portion of the mitochondrial 16S ribosomal RNA gene by polymerase chain reaction (PCR), using evolutionarily conserved primers, from a selection of species representative of the genera Rhodnius, Triatoma and Panstrongylus. Amplification products were subsequently screened for sequence variation using single strand conformational polymorphism analysis (SSCP) and also subjected to direct sequencing. Single strand conformational polymorphism analysis could detect variation within and between genera; intraspecific variation was also detected and SSCP profiles appear to be useful for identification purposes at the inter- and intraspecific levels. A 290 bp multiple alignment of 15 sequences obtained from nine species was generated, phylogenetic inference subsequently used three methods; a distance estimate, maximum parsimony and maximum likelihood. This 16S region exhibited considerable variation which ranged from intergeneric to intraspecific levels. Phylogenies from these three methods of inference were in broad agreement. Triatoma and Panstrongylus were more closely related to each other than either was to Rhodnius, in keeping with the current taxonomic appraisal.

Identification of a novel HLA-DRB1*04:94N allele by polymerase chain reaction sequence-based typing

2011 ◽  
Vol 78 (3) ◽  
pp. 226-227 ◽  
Author(s):  
F.-M. Zhu ◽  
W. Wang ◽  
W. Zhang ◽  
H.-J. Lv ◽  
L.-X. Yan
Keyword(s):  
Polymerase Chain Reaction ◽  
Reaction Sequence ◽  
Chain Reaction ◽  
Polymerase Chain

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.

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