Background Small RNA sequencing (RNA‐seq) of biofluids is challenging due to the relative scarcity of microRNAs (miRNAs), limited sample volumes, and the lack of a gold standard isolation method. Additionally, few comparisons exist for the RNA isolation and sequencing methods ...
Read More »Incorporation of unique molecular identifiers (UMIs) in TruSeq adapters improves the accuracy of quantitative sequencing
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. ...
Read More »The impact of amplification on differential expression analyses by RNA-Seq
Currently quantitative RNA-Seq methods are pushed to work with increasingly small starting amounts of RNA that require PCR amplification to generate libraries. However, it is unclear how much noise or bias amplification introduces and how this effects precision and accuracy ...
Read More »Low-cost, low-input RNA-seq protocols perform nearly as well as high-input protocols
Recently, a number of protocols extending RNA-sequencing to the single-cell regime have been published. However, we were concerned that the additional steps to deal with such minute quantities of input sample would introduce serious biases that would make analysis of ...
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