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....
| Main Authors: | , , |
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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 |
| Summary: | 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. |
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Optimization of the SSCP-HD technique for the detection of polymorphisms on exon 3 of the ADIPOQ gene