Ciencia e Ingeniería en Alimentos y Biotecnología

URI permanente para esta comunidadhttp://repositorio.uta.edu.ec/handle/123456789/412

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2022 - 20262

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Fecha de inicio: 2022Tema: search.filters.subject.GENÉTICA

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  • Miniatura
    Item type:Ítem,
    Desarrollo de formulaciones locales de tampón de carga y marcador de peso molecular de 100 pb para electroforesis en gel de agarosa
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07)
    Avalos Chávez, Angie Valeria
    ;
    Calero Cáceres, William Ricardo
    Irregular access to molecular biology reagents limits genetic research in Latin America due to high importation costs, dependence on international suppliers, and regulatory constraints that hinder scientific continuity in resource-limited settings. Natural pigments, including anthocyanins, betalains, chlorophylls, curcumin, capsanthin, and norbixin, as well as food-grade synthetic dyes, were evaluated through electrophoretic screening. Loading buffer formulations were prepared by combining two base buffer systems (60 mM Tris-HCl, pH 7.6, and 6X TE buffer) with different dye concentrations. Chemical stability was monitored by UV–Vis spectrophotometry over 120 days under three storage conditions. The molecular weight marker was generated from PCR products amplified from the rpoB gene of E. coli. The results showed that Brilliant Blue FCF (E133) and Tartrazine (E102) were colorants that showed compatibility with electrophoretic conditions. The T7 formulation, consisting of Tris-HCl without EDTA and a high concentration of both dyes, stored at 20 Celcius degree, exhibited the highest spectral and electrophoretic stability throughout the evaluation period. The developed molecular marker produced a reproducible banding pattern ranging from 400 to 2000 bp, with performance comparable to that of commercial references. These findings demonstrate that the local production of functional electrophoretic reagents is technically feasible using accessible and low-cost materials. This study provides a reproducible in-house methodology for the generation of electrophoresis reagents, with the potential to reduce the technological gap in laboratories located in low- and middle-income countries.
  • Miniatura
    Item type:Ítem,
    Diseño y validación in silico de primers para la amplificación del Gen GSTM
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-03)
    Guevara Guevara, Kevin Joel
    ;
    Galarza Galarza, Cristian Fernando
    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.