Detailed analysis of HTT repeat elements in human blood using targeted amplification-free long-read sequencing.

ABSTRACT: Amplification of DNA is required as a mandatory step during library preparation in most targeted sequencing protocols. This can be a critical limitation when targeting regions that are highly repetitive or with extreme guanine-cytosine (GC) content, including repeat expansions associated with human disease. Here, we used an amplification-free protocol for targeted enrichment utilizing the CRISPR/Cas9 system (No-Amp Targeted sequencing) in combination with single molecule, real-time (SMRT) sequencing for studying repeat elements in the huntingtin (HTT) gene, where an expanded CAG repeat is causative for Huntington disease. We also developed a robust data analysis pipeline for repeat element analysis that is independent of alignment of reads to a reference genome. The method was applied to 1

SUBMITTER: Hoijer I 

PROVIDER: S-EPMC6175010 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature