| abstract |
Methods for preparing sequencing libraries based on a plurality of RNA molecules which are tagged and amplified by tagging the molecule with an oligonucleotide hybridising to a polyC tail introduced by the terminal transferase activity of the reverse transcriptase, e.g. MMLV RT, and ligating the oligonucleotide to the RNA molecule using a ligase, e.g. T4 RNA ligase, and producing cDNA molecules based on mRNA and strand displacing reverse transcriptases, e.g.g MMLV RT, and producing a second cDNA strand in order to produce a dsDNA library for sequencing. In some embodiments, the subject methods comprise sequencing at least a portion of a sequencing library to obtain sequencing data or sequence reads from a test sample (e.g., a biological sample from a subject). |