polyA: The 15-40 nt fraction derived from 1 μg small RNA (<200 nt) was polyadenylated, purified and treated with periodic acid (as in 2) for 3` termination of the poly(A) tail. After ligation of the 5` linker, the >70 nt fraction was excised from 10% denaturing polyacrylamide gel (for replicate 1) or this step was omitted (replicate 2). Reverse-transcription was performed with the poly(A) RT primer, followed by 15 cycli of PCR (replicate 1) or 1/10 of the RT reaction was amplified by 17 cycles of PCR (replicate 2).
Library strategy
RNA-Seq
Library source
transcriptomic
Library selection
size fractionation
Instrument model
AB SOLiD System 2.0
Description
none
Data processing
Sequencing reads were mapped to the known miRNA and miRNA* species in miRBase v11.0 using SHRiMP. Reads with 3 or less colorspace errors were included in the further analysis.
