Next-generation sequencing (NGS) provides an unprecedented opportunity to assess genetic variation underlying human disease. Here, we compared two NGS approaches for diagnostic sequencing in inherited arrhythmia syndromes. We compared PCR-based target enrichment and long-read sequencing (PCR-LR) with in-solution hybridization-based enrichment and short-read sequencing (Hyb-SR). The PCR-LR assay...

Indexing individual template molecules with a unique identifier (UID) before PCR and deep sequencing is promising for detecting low frequency mutations, as true mutations could be distinguished from PCR errors or sequencing errors based on consensus among reads sharing same index. In an effort to develop a robust assay to detect from urine low-abundant bladder cancer cells carrying well-documen...

Quantitative analysis of next-generation sequencing (NGS) data requires discriminating duplicate reads generated by PCR from identical molecules that are of unique origin. Typically, PCR duplicates are identified as sequence reads that align to the same genomic coordinates using reference-based alignment. However, identical molecules can be independently generated during library preparation. Mi...

PCR permits the exponential and sequence-specific amplification of DNA, even from minute starting quantities. PCR is a fundamental step in preparing DNA samples for high-throughput sequencing. However, there are errors associated with PCR-mediated amplification. Here we examine the effects of four important sources of error-bias, stochasticity, template switches and polymerase errors-on sequenc...

a. We recommend that the standard methodology for genetic diagnosis of autosomal dominant polycystic kidney disease is polymerase chain reaction (PCR) amplification (including long-range PCR for the first 33 exons of PKD1) followed by Sanger sequencing (1A) or next-generation sequencing where available (1D). b. We suggest that individuals with a clinical diagnosis of autosomal dominant polycyst...

Background and Aims: lamivudine is amongst the antiviral for drug chronic hepatitis B treatment. During therapy with lamivudine, variants may emerge with YMDD mutation in the reverse transcriptase (RT) region of polymerase gene. This mutation might have a role in drug resistant for HBV. Materials and Methods: HBV DNA extraction from serum sample of 88 patients, were subjected to nested PCR for ...

Mutations in the gene encoding filaggrin (FLG) have been identified as cause of ichthyosis vulgaris (IV) and shown to be major predisposing factors for atopic dermatitis. The profilaggrin protein consists 10-12 monomer repeats arranged tandem, which arise from an ultra-long sequence exon 3 comprised near-indistinguishable repeats, demonstrating intragenic copy number variations (CNVs). As such,...

  Target modification and reduced drug accumulation are the main resistance mechanisms against fluoroquinolone antibiotics in Pseudomonas aeruginosa. We performed a genotypic characterization of three major Mex multidrug efflux pumps (MexAB-OprM, MexXY-OprM and MexCD-OprJ) in ciprofloxacin resistant clinical isolates of P. aeru­ginosa, collected from Tehran, Iran this was followed by sequencin...

Amplicon (targeted) sequencing by massively parallel sequencing (PCR-MPS) is a potential method for use in forensic DNA analyses. In this application, PCR-MPS may supplement or replace other instrumental analysis methods such as capillary electrophoresis and Sanger sequencing for STR and mitochondrial DNA typing, respectively. PCR-MPS also may enable the expansion of forensic DNA analysis metho...