Abstract
Combining next-generation sequencing with capture of sequences from a relevant subset of a transcriptome produces an enhanced view of this subset
Highlights
In recent years, a technologic revolution has shifted DNA sequencing from traditional Sanger methods to "next-generation" sequencing
Results cDNA hybrid selection To develop a targeted RNA-Seq method, we created a complex pool of biotinylated oligonucleotide probes for cancer-related transcripts and used them to capture cDNAs from a library prepared for Illumina sequencing
A cDNA library for Illumina sequencing was constructed from the K-562 chronic myeloid leukemia (CML) cell line
Summary
A technologic revolution has shifted DNA sequencing from traditional Sanger methods to "next-generation" sequencing (see review [1]). Applying these new sequencing methods to cDNA libraries, termed RNA-Seq, generates a wealth of information beyond that obtained from sequencing genomic DNA (see review [2]). Chromosomal rearrangements can be detected by next-generation sequencing of genomic DNA [9,10], RNA-Seq is a powerful tool to identify those rearrangements that lead to chimeric transcripts and are more likely to have functional consequences in cancer [3,11]
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