Removing bias against short sequences enables northern blotting to better complement RNA-seq for the study of small RNAs.

ABSTRACT: Changes in small non-coding RNAs such as micro RNAs (miRNAs) can serve as indicators of disease and can be measured using next-generation sequencing of RNA (RNA-seq). Here, we highlight the need for approaches that complement RNA-seq, discover that northern blotting of small RNAs is biased against short sequences and develop a protocol that removes this bias. We found that multiple small RNA-seq datasets from the worm Caenorhabditis elegans had shorter forms of miRNAs that appear to be degradation products that arose during the preparatory steps required for RNA-seq. When using northern blotting during these studies, we discovered that miRNA-length probes can have ∼1000-fold bias against detecting even synthetic sequences that are 8 nt shorter. By using shorter probes and by performing hyb

SUBMITTER: Choi YS 

PROVIDER: S-EPMC5449620 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature