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

Hum. Mutat.. 2018-09; 
HöijerIda,TsaiYu-Chih,ClarkTyson A,KotturiPaul,DahlNiklas,StattinEva-Lena,BondesonMarie-Louise,FeukLars,GyllenstenUlf,Ameur
Products/Services Used Details Operation
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

摘要

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

关键词

HTT,Huntington disease,No-Amp Targeted sequencing,SMRT sequencing,amplification-free sequencing,repeat expansion,somatic mosaicism,targeted enrichment,targeted sequen