Current major barriers for using next-generation sequencing (NGS) technologies 
in genetic mutation screening on an epidemiological scale appear to be the high 
accuracy demanded by clinical applications and high per-sample cost. How to 
achieve high efficiency in enriching targeted disease genes while keeping a low 
cost/sample is a key technical hurdle to overcome. We validated a 
cDNA-probe-based approach for capturing exons of a group of genes known to cause 
deafness. Polymerase chain reaction amplicons were made from cDNA clones of the 
targeted genes and used as bait probes in hybridization for capturing human 
genomic DNA (gDNA) fragments. The cDNA library containing the clones of targeted 
genes provided a readily available, low-cost, and regenerable source for 
producing capture probes with standard molecular biology equipment. Captured 
gDNA fragments by our method were sequenced by the Illumina NGS platform. 
Results demonstrated that targeted exons captured by our approach achieved 
specificity, multiplexicity, uniformity, and depth of coverage suitable for 
accurate sequencing applications by the NGS systems. Reliable genotype calls for 
various homozygous and heterozygous mutations were achieved. The results were 
confirmed independently by conventional Sanger sequencing. The method validated 
here could be readily expanded to include all-known deafness genes for 
applications such as genetic hearing screening in newborns. The high coverage 
depth and cost benefits of the cDNA-probe-based exon capture approach may also 
facilitate widespread applications in clinical practices beyond screening 
mutations in deafness genes.