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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.