Barcoding bias in high-throughput multiplex sequencing of miRNA

• DOI: 10.1101/gr.121715.111
• Publication Date: 2011-07-13T04:59:43Z
• Authors: Shahar Alon; Francois Vigneault; Seda Eminaga; Danos C. Christod...; Jonathan G. Seidman; George M. Church; Eli Eisenberg
• Keywords: Barcode; Multiplex; Adapter (computing)

Second-generation sequencing is gradually becoming the method of choice for miRNA detection and expression profiling. Given relatively small number miRNAs improvements in DNA technology, studying profiles multiple samples a single flow cell lane becomes feasible. Multiplexing strategies require marking each library with barcode. Here we report that barcodes introduced through adapter ligation confer significant bias on profiles. This much higher than expected Poisson noise masks differences between libraries. can be eliminated by adding during PCR amplification The accuracy measurement multiplexed experiments function sample number.