Development of a multi-dimensional pooling approach for scanning rare mutations in deep sequencing experiments and application on familial hypercholesterolemia patient group

Tests based on DNA sequencing analysis have only limited application on screening of genetic diseases in masses, due to their high costs. The pooling approach especially used in disease screening programs, due to its cost reduction benefits. The combination of Next Generation Sequencing (NGS) technology and pooling approach can be used in screening of genetic diseases through the populations as it offers a solution to the high cost problem. In this thesis, different 2 and 3 dimensional pooling approaches are tested without compromising the ease of use. Then, 256 FH patients and 64 control samples were screened through their 6 genes by using NGS technology and 2 dimensional 8x8 pooling approach, which is selected through testing different pooling approaches. Among different pooling approaches tested, 2 dimensional 8x8 and 16x16 pooling strategies are evaluated as successful and 24x24 pooling is evaluated as partially successful. The higher pooling approaches are evaluated as unsuccessful. Among 320 samples screened, 89 different genotypes are detected from 40 different mutations in 75 of the samples.