dna sequencing sequencher heterozygous mutations

Hartman P., Beckman K., Silverstein K., Yohe S., Schomaker M., Henzler C., Onsongo G., Lam H.C., Munro S., Daniel J., et al. Our results indicate that targeted double-stranded cDNA sequencing-based phase analysis is useful for determining compound heterozygous mutations and confers information on allelic expression.Īllelic expression compound heterozygous mutation next-generation sequencing phase analysis targeted double-stranded cDNA sequencing. Furthermore, targeted double-stranded cDNA sequencing-based phase analysis detected the differential allelic expression, suggesting that the mutation (chr13:51958362 G>GG) caused downregulation of expression by nonsense-mediated mRNA decay. Trio analysis showed that the mutation (chr13:51964900 C>T) derived from the father and the other mutation from the mother, validating that the mutations are in trans composition. Targeted double-stranded cDNA sequencing-based phase analysis was performed using direct adapter ligation library preparation and paired-end sequencing, and we elucidated they are in trans compound heterozygous mutations. The distance between the two mutations was 6.5 kb in genomic DNA but 464 bp in mRNA. Here, we performed a genetic analysis on a patient with Wilson disease, and we detected two heterozygous mutations chr13:51958362 G>GG (NM_000053.4:c.2304dup r.2304dup p.Met769HisfsTer26) and chr13:51964900 C>T (NM_000053.4:c.1841G>A r.1841g>a p.Gly614Asp) in the causative gene ATP7B. However, conventional phase analysis is limited because of the large target size of genomic DNA. Because only in trans compound heterozygous mutations have been implicated in autosomal recessive diseases, it is important to distinguish them for clinical diagnosis. There are two combinations of heterozygous mutation, i.e., in trans, which carries mutations on different alleles, and in cis, which carries mutations on the same allele.

Dna sequencing sequencher heterozygous mutations