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Detection of mutations and intragenic rearrangements in BRCA1 and BRCA2 genes by sequencing and MLPA method
PECHOVÁ, Kristýna
In my bachelor thesis I dealt with the detection of mutations in BRCA genes and their effect on the development of breast cancer. Nowadays, this topic is very important, because breast cancer is the most common cancer among women in the Czech Republic (Weinberger a Zikán, 2016). In the theoretical part I focused on breast cancer, particularly on its diagnosis and consequent treatment. I also dedicated genetic counselling and examination, which is very important for the diagnosis of hereditary forms of cancer. I mentioned basic information about BRCA genes, and their protein products and I summarized the issue of mutations in these genes and their relationship to cancer, particularly breast and ovarian. In the practical part, I focused on the examination of selected areas of BRCA genes by the Sanger sequencing method. At present, BRCA genes are examined by the NGS method, because it allows for the analysis of all exons of one particular gene at once, but this testing wasn't possible to realize in my laboratory conditions, therefore I have chosen the method of Sanger sequencing, which does, however, have certain capacity limitations. By using the PCR method, I prepared samples for testing by Sanger sequencing. This testing was conducted by the firm Genseq s.r.o. In the last part of my thesis I processed the obtained sequences and I evaluated the results. I examined a total of 20 anonymized samples using the sequencing method. Mutation was present in 4 samples of the total amount - one was pathogenic, one benign and two of uncertain significance.
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