Modban: The original protocol, as described by Lau et al. 2001, was followed. We used as input the 15-40 nt fraction of 1 μg of small RNA isolated using the miRVana kit. A preadenylated modban adapter was ligated to the to the RNA in the absence of ATP. The 30-50 nt fraction was excised from a 15% denaturing polyacrylamide gel. After elution in 0.3M NaCL and EtOH precipitation, the 5` linker was ligated in the presence of ATP. The 70-90 nt fraction was gel-excised from a 10% denaturing polyacrylamide gel, eluted and precipitated. Reverse-transcription was performed with the modban RT primer and 15 cycles of PCR were performed to amplify the library.
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.
