Diseño y validación in silico de primers para la amplificación del Gen GSTM

Abstract

Lung cancer is a disease that affects a large part of the population, there are several reasons why it develops, one of them involves the GSTM1 gene, for this reason the possible relationship with cancer was consulted through the databases Malacards and Harmonizome, using the NM_000561.4 sequence for primer design. The region taken for the design of the primers comprised between exon 5 and exon 7, since in this there is a SNP related to lung cancer, a substitution of a C for a G in position 534, resulting in the GSTM1 A and GSTM1 B alleles containing a lysine and an asparagine respectively at position 172, a third GSTM1 0 allele called the null allele generated by a homozygous deletion, which leads to susceptibility to lung cancer. Hence the importance of design for both diagnostic and research use. The design was carried out in the Primer-Blast and Primer3Plus programs with default parameters and another design modifying them, mainly the concentrations of divalent cations and dNTPs, the rest of the configurations were modified according to the purpose of the primers. Once the set of primers was obtained, compatibility was analyzed using the MultiPLX 2.1 program and the melting curves were obtained using the uMELT software. Finally, the in silico validation was carried out with the Oligoanalyzer and Beacon designer free edition programs.