The set of exons selected for this mutation scanning analysis were drawn from cancer genes frequently mutated (PIK3CA, ERBB2, KRAS, TP53, EGFR, BRAF, GATA3, and FGFR3) in published reports, with a particular emphasis on breast, ovarian and endometrium cancers [5], [9], [27], [28], [29], [30], [31], [32].
The aim of this study was to assess the sensitivity and specificity of an inexpensive HRM analysis platform for mutation scanning of single-base variation in a range of tumor samples: frozen pediatric small rounded blue-cell tumors and paraffin-embedded tumors from breast, endometrium and ovarian cancers.
For MRN amplicons harboring a SNP s) with frequency ≥1% in either dbSNP or initial amplicon testing, we applied a simultaneous mutation scanning and genotyping approach using HRM analysis to improve the sensitivity and the efficiency of the mutation screening, as described previously [ 24, 25].
For CHEK2 amplicons harboring SNPs with a frequency ≥1% in either the Single Nucleotide Polymorphism Database (dbSNP) [ 33] or initial amplicon testing, we applied a simultaneous mutation scanning and genotyping approach using HRM curve analysis to improve the sensitivity and efficiency of the mutation screening [ 34].
We carried out mutation screening of the nine exons and intron/exon boundaries of RAD51C by high resolution melting (HRM) analysis, a rapid closed-tube mutation scanning method with high sensitivity and specificity.
Linkage analysis, positional cloning, candidate gene mutation scanning and genome-wide association study approaches have all contributed significantly to our understanding of the underlying genetic architecture of breast cancer.
HRM analysis is the next generation of mutation scanning technology and can be cost-effective, requiring less time for results when compared to other scanning techniques.
Molecular analysis for OI includes sequencing or mutation scanning and deletion/duplication testing for COL1A1 and COL1A2, followed by sequencing of recessive OI-related genes (see Table 1).
We previously validated mutation scanning for BRCA1 and 2 using high-resolution melting curve analysis (HRMCA).
High-resolution melting (HRM) analysis is a very effective method for genotyping and mutation scanning that is usually performed just after PCR amplification (the "closed tube" format).
High-resolution melting curve analysis (hrMCA) has recently been developed as a post-PCR mutation scanning method which enables simple, rapid, cost-effective, and highly sensitive mutation screening of large genes.
