We constructed a cDNA library from small RNAs purified from Q column fractions of testes extracts. Briefly, gel-purified small RNAs from the column fractions were ligated with adaptor oligos on both 3' and 5' ends of the small RNAs. Reverse-transcription and PCR amplification was then performed on the ligated small RNAs to yield a cDNA library. This cDNA library was finally converted by PCR into a single-stranded DNA library compatible for coupling to beads on the 454 Life Sciences Genome Sequencer 20 System. Reads and sequences of the small RNAs were processed from linker sequences, examined for matches to the mouse genome, and tallied.
Description
Sequences were analyzed and annotated for matches to the mouse genome. See Supplemental Online Materials from Lau et al. Science. 2006 Jun 15; [Epub ahead of print] PMID: 16778019 for detailed protocol on data processing.
