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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
Oliveira de Miranda, Vitor Fernandes [1] ; Martins, Vanderlei Geraldo [2, 3] ; Furlan, Antonio [4] ; Bacci, Jr., Mauricio [2]
Total Authors: 4
Affiliation:
[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
Total Affiliations: 4
Document type: Journal article
Source: Brazilian Archives of Biology and Technology; v. 53, n. 1, p. 141-152, JAN-FEB 2010.
Web of Science Citations: 6
Abstract

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)