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Gene Synthesis> | … NA20248, NA20251 and NA20252). HEK 293 genomic DNA, used for No-Amp Targeted sequencing, was purchased from Genscript. Fragment analysis of HTT repeat expansions All samples and controls were subjected to PCR with primers designed to target the CAG … | Get A Quote |
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... More