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Table 1 Summary of the principal characteristics of the RNA-Seq library preparation kits evaluated in this study

From: Comparison of library preparation methods reveals their impact on interpretation of metatranscriptomic data

  TruSeq stranded Encore complete Ovation RNA-Seq V2 SMARTer stranded
RNA input (minimal requirement according to the manufacturer) 100 ng depleted RNA 100 ng total RNA 0,5 ng depleted RNA 1 ng depleted RNA
Minimum quality FFPE RIN >7 FFPE FFPE
rRNA depletion required Yes No Yes Yes
cDNA synthesis Random primers Selective priming Random and oligo(dT) primers Random primers
Fragmentation method RNA by divalent cations + heat cDNA by Covaris shearing cDNA by Covaris shearing RNA by heat
Strand selection Yes Yes No Yes
Library preparation method and reagents Included Included Not included Included
Multiplex capacity 96-plex 16-plex according to the library preparation method chosen 12-plex
Experiment duration 6 hours 7 hours 4.5 hours for cDNA synthesis 4.5 hours
+ time for library preparation
  1. FFPE: Formalin-fixed, paraffin-embedded tissue.
  2. RIN: RNA integrity number.