Plant or Fungal Sequences? An Alternative Optimized PCR Protocol to Avoid ITS (nrDNA) Misamplification

Autor(es)

Oliveira de Miranda, Vitor Fernandes ^[1] ; Martins, Vanderlei Geraldo ^{[2, 3]} ; Furlan, Antonio ^[4] ; Bacci, Jr., Mauricio ^[2]

Afiliação do(s) autor(es)

^[1] Univ Mogi Das Cruzes, Lab Sistemat Vegetal & Herbarium Mogiense, BR-08780911 Mogi Das Cruzes, SP - Brazil

^[2] Univ Estadual Paulista, Ctr Estudos Insetos Sociais, BR-13506900 Rio Claro, SP - Brazil

^[3] Univ Estadual Paulista, Dept Fonoaudiol, Marilia, SP - Brazil

^[4] Univ Estadual Paulista, Dept Bot, BR-13506900 Rio Claro, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Brazilian Archives of Biology and Technology; v. 53, n. 1, p. 141-152, JAN-FEB 2010.
• Citações Web of Science: 6

Resumo

The nuclear ribosomal DNA internal transcribed spacers (ITS1 and ITS2) from leaves of Drosera (Droseraceae) were amplified using ``universal{''} primers. The analysis of the products demonstrated most samples were a molecular mixture as a result of unsuccessful and non-specific amplifications. Among the obtained sequences, two were from Basidiomycota fungi. Homologous sequences of Basidiomycota were obtained from GenBank database and added to a data set with sequences from Drosera leaves. Parsimony analysis demonstrated that one sequence was amplified from an Ustilaginomycetes fungus, and another from a Heterobasidiomycetes. Possibly these fungi were associated to leaves of Drosera, and not because of samples contamination. In order to provide optimization and a better specificity of PCR (polymerase chain reaction), a very successful method was demonstrated using dimethyl sulfoxide (DMSO) and bovine serum albumin (BSA) in reactions. (AU)

• Processo FAPESP: 00/05098-9 - Análise filogenética de espécies americanas e africanas do gênero Drosera (Droseraceae) utilizando caracteres morfológicos e moleculares
• Beneficiário: Vitor Fernandes Oliveira de Miranda
• Modalidade de apoio: Bolsas no Brasil - Mestrado