Authors: Maurício Papa de Arruda Evonnildo Costa Gonçalves Maria Paula Cruz Schneider Artur Luiz da Costa da Silva Eliana MorielleVersute
Publish Date: 2009/07/30
Volume: 37, Issue: 4, Pages: 2031-2036
Abstract
We proposed a modification the procedure of genotyping based in labeled universal primer and tailed primer In the standard protocol three primers are used in the same PCR reaction a forward primer with tail added at the 5′ end of the identical sequence to labeled universal primer with dyefluorescent and a reverse primer Unfortunately the choice of a labeled primer characterized by a large number of complementary sequences in target genomes which is more probable in larger genomes result in unspecific amplifications false positive can cause absence or decrease amplification of the locus of interest and also false interpretation of the analysis However identification of possible homologies between the primer chosen for labelling and the genome is rarely possible from the available DNA data bases In our approach cycling is interrupted for the addition of the labeled primer only during the final cycles thus minimizing unspecific amplification and competition between primers resulting in the more fidelity amplification of the target regions
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