Chronic lymphocytic leukemia (CLL) represents a prototype disease in which TP53 gene defects lead to inferior prognosis. Here, we present two distinct methodologies which can be used to identify TP53 mutations in CLL patients; both protocols are primarily intended for research purposes. The functional analysis of separated alleles in yeast (FASAY) can be flexibly adapted to a variable number of samples and provides an immediate functional readout of identified mutations. Amplicon-based next-generation sequencing then allows for a high throughput and accurately detects subclonal TP53 variants (sensitivity <1% of mutated cells).

• MeSH
• Alleles MeSH
• Leukemia, Lymphocytic, Chronic, B-Cell blood   genetics   pathology   MeSH
• Humans MeSH
• Mutation MeSH
• DNA Mutational Analysis instrumentation   methods   MeSH
• Neoplastic Cells, Circulating pathology   MeSH
• Tumor Suppressor Protein p53 genetics   MeSH
• Genes, Reporter genetics   MeSH
• Saccharomyces cerevisiae genetics   MeSH
• Transfection instrumentation   methods   MeSH
• High-Throughput Nucleotide Sequencing instrumentation   methods   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH