Amplification optimization through PCR for region 1 promoter of gen shp-1 in bisulfite treated region
Changes in methylation patterns may contribute to cancer development. Recent studies have shown that shp-1 expression is diminished in prostate cancer. Accordingly, it is relevant to evaluate the methylation pattern of its promoters. Bisulfite sequencing is the reference technique for these studies....
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| Format: | Online |
| Language: | Spanish |
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Facultad de Ciencias Exactas, Químicas y Naturales
2009
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| Subjects: | |
| Online Access: | https://www.fceqyn.unam.edu.ar/recyt/index.php/recyt/article/view/529 |
| _version_ | 1814554273541783552 |
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| author | Giorgio, E. Martín Chaneton, Bárbara J. Zapata, Pedro D. |
| author_facet | Giorgio, E. Martín Chaneton, Bárbara J. Zapata, Pedro D. |
| author_sort | Giorgio, E. Martín |
| collection | Revista de Ciencia y Tecnologia RECyT |
| description | Changes in methylation patterns may contribute to cancer development. Recent studies have shown that shp-1 expression is diminished in prostate cancer. Accordingly, it is relevant to evaluate the methylation pattern of its promoters. Bisulfite sequencing is the reference technique for these studies. The aim of this paper was the standardization of the conditions for shp-1 gene Promoter 1 amplification before bisulfite treatment. Two tissues with differential expression of shp-1 were investigated: buccal epithelial cells and blood. We analyzed different combinations of PCR component with primers that hybridized in CpG island free zones. The application of 3 segments of decreased hybridization temperature allowed us to obtain 500pb predicted amplicons for this fragment of promoter 1. |
| format | Online |
| id | ojs-article-529 |
| institution | Facultad de Ciencias Exactas Quimicas y Naturales |
| language | Spanish |
| publishDate | 2009 |
| publisher | Facultad de Ciencias Exactas, Químicas y Naturales |
| record_format | ojs |
| spelling | ojs-article-5292020-03-06T10:52:29Z Amplification optimization through PCR for region 1 promoter of gen shp-1 in bisulfite treated region Optimización de la amplificación mediante PCR de la región promotora 1 del gen shp-1 luego de su tratamiento con bisulfito Giorgio, E. Martín Chaneton, Bárbara J. Zapata, Pedro D. epygenome bisulfite treatment PCR shp-1 epigenoma tratamiento con bisulfito PCR shp-1 Changes in methylation patterns may contribute to cancer development. Recent studies have shown that shp-1 expression is diminished in prostate cancer. Accordingly, it is relevant to evaluate the methylation pattern of its promoters. Bisulfite sequencing is the reference technique for these studies. The aim of this paper was the standardization of the conditions for shp-1 gene Promoter 1 amplification before bisulfite treatment. Two tissues with differential expression of shp-1 were investigated: buccal epithelial cells and blood. We analyzed different combinations of PCR component with primers that hybridized in CpG island free zones. The application of 3 segments of decreased hybridization temperature allowed us to obtain 500pb predicted amplicons for this fragment of promoter 1. Las alteraciones en los patrones de metilación de algunos genes pueden contribuir al desarrollo del cáncer. Existen evidencias de cambios en los niveles de expresión de SHP-1 en próstata tumoral, siendo fundamental evaluar el estado de metilación de sus promotores. La secuenciación bisulfito es la técnica de elección para establecer el patrón de metilación. El objetivo de este trabajo fue estandarizar las condiciones para la amplificación del Promotor 1 para el gen shp-1 luego del tratamiento con bisulfito. Se trabajó con 2 tejidos de expresión diferencial para SHP-1: mucosa yugal y tejido sanguíneo. A partir de cebadores que hibridan en sitios libres de islas CpG del promotor 1 del gen shp-1 tratado con bisulfito de sodio se analizaron distintas combinaciones de los componentes de PCR aplicándose distintas estrategias de ciclado. Un esquema de ciclado con 3 bloques de temperatura de hibridación decreciente permitió la obtención del amplicón de 500pb coincidente con el esperado para el fragmento del promotor analizado. Facultad de Ciencias Exactas, Químicas y Naturales 2009-12-20 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://www.fceqyn.unam.edu.ar/recyt/index.php/recyt/article/view/529 Argentine Journal of Science and Technology; Vol. 12 No. 1 (2009); 4-8 Revista de Ciencia y Tecnología; Vol. 12 Núm. 1 (2009); 4-8 1851-7587 0329-8922 spa https://www.fceqyn.unam.edu.ar/recyt/index.php/recyt/article/view/529/440 Derechos de autor 2009 Revista de Ciencia y Tecnología |
| spellingShingle | epygenome bisulfite treatment PCR shp-1 epigenoma tratamiento con bisulfito PCR shp-1 Giorgio, E. Martín Chaneton, Bárbara J. Zapata, Pedro D. Amplification optimization through PCR for region 1 promoter of gen shp-1 in bisulfite treated region |
| title | Amplification optimization through PCR for region 1 promoter of gen shp-1 in bisulfite treated region |
| title_alt | Optimización de la amplificación mediante PCR de la región promotora 1 del gen shp-1 luego de su tratamiento con bisulfito |
| title_full | Amplification optimization through PCR for region 1 promoter of gen shp-1 in bisulfite treated region |
| title_fullStr | Amplification optimization through PCR for region 1 promoter of gen shp-1 in bisulfite treated region |
| title_full_unstemmed | Amplification optimization through PCR for region 1 promoter of gen shp-1 in bisulfite treated region |
| title_short | Amplification optimization through PCR for region 1 promoter of gen shp-1 in bisulfite treated region |
| title_sort | amplification optimization through pcr for region 1 promoter of gen shp 1 in bisulfite treated region |
| topic | epygenome bisulfite treatment PCR shp-1 epigenoma tratamiento con bisulfito PCR shp-1 |
| topic_facet | epygenome bisulfite treatment PCR shp-1 epigenoma tratamiento con bisulfito PCR shp-1 |
| url | https://www.fceqyn.unam.edu.ar/recyt/index.php/recyt/article/view/529 |
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Optimization of the SSCP-HD technique for the detection of polymorphisms on exon 3 of the ADIPOQ gene