Genetics and Molecular Research 16 (2): gmr16029647
Development of TRAP primers for
Ricinus communis L.
K.S. Simões, S.A. Silva, E.L. Machado and H.S. Brasileiro
Núcleo de Melhoramento Genético e Biotecnologia,
Centro de Ciências Agrárias Ambientais e Biológicas,
Universidade Federal do Recôncavo da Bahia, Cruz das Almas, BA, Brasil
Corresponding author: K.S. Simões
E-mail: karinesimoes01@hotmail.com
Genet. Mol. Res. 16 (2): gmr16029647
Received February 16, 2017
Accepted March 16, 2017
Published April 13, 2017
DOI http://dx.doi.org/10.4238/gmr16029647
Copyright © 2017 The Authors. This is an open-access article distributed under the terms of
the Creative Commons Attribution ShareAlike (CC BY-SA) 4.0 License.
ABSTRACT. The objective of this article was to develop TRAP (target
region amplifcation polymorphism) primers for castor bean, with the
goal of making functional markers available for genetic studies about
the species. To do this, oligonucleotides were designed based on
ESTs, obtained from the NCBI (National Center for Biotechnology
Information) databank, which code enzymes involved in metabolic
routes of fatty acid synthesis, ricin synthesis, and resistance to castor
bean pathogens. The forward primers were designed with the help of
the Primer3 software and, for the reverse, six arbitrary primers were
used. To standardize the amplifcation reactions, the following criteria
were used to select the primers: sizes between 18 and 20 bp, guanine/
cytosine (GC) in the range of 40 to 60%, and average annealing
temperature between 55° and 62°C. The design quality of the primers
was verifed using the Net Primer application. Fifty-six primers were
designed, which had an average GC percentage of 53.2%. A total of
336 combinations were obtained using the 56 fxed and 6 arbitrary
primers. Based on polymerase chain reaction, 330 combinations (89%)
presented good amplifcation patterns for the genomic DNA of castor