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Table 1 Comparison of library construction protocols

From: Strand-specific libraries for high throughput RNA sequencing (RNA-Seq) prepared without poly(A) selection

Method Time (days) Total steps Reagent cost ($) Starting RNA (ng) Use for small RNA Kits available Barcoding described
This protocol 2 9 38 50 No No Yes
RNA ligation 8 19 250 1200 Yes No No
Illumina RNA ligation 5 16 240 100 Yes Partially No
Illumina RNA ligation-SPRI 4 12 220 100 Yes Partially No
SMART 5 8 80 100 Yes No No
Hybrid 5 13 90 500 Yes No No
NNSR 4 9 90 500 Unclear No No
NNSR, no Actinomycin D 4 9 90 500 Unclear No No
Bisulfite ‘S’ 6 19 540 1000 No Mostly No
Bisulfite ‘H’ 6 19 540 1000 No Mostly No
dUTP 5 17 430 200 No Mostly No
dUTP oligo(dT) 5 17 440 200 No Mostly No
  1. Data for other methods from Levin [12]. Values only include steps after rRNA removal (this protocol) or poly(A) selection. dUTP: deoxyuridine triphosphate; NNSR: not not so random; SMART: switching mechanism at 5’ end of RNA transcript; SPRI, solid phase reversible immobilization.