ScriptSeq™ mRNA-Seq Library Preperation Kit (Illumina-compatible)

ScriptSeq™ mRNA-Seq Library Preperation Kit (Illumina-compatible)




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The ScriptSeq™ mRNA-Seq Library Preparation Kit (Illumina-compatible)* uses a unique terminal-tagging technology* that simplifies the preparation of directional, paired-end libraries from rRNA-depleted or poly(A)-enriched RNA (Fig. 1). The kit is suitable for any animal, plant, or bacterial species, and enables library preparation in about 3 hours.

The ScriptSeq™ mRNA-Seq Library Preparation Kit (Illumina-compatible):

  • Enables strand-specific adaptor-tagging for directional paired-end sequencing using standard Illumina® sequencing primers.
  • Produces cluster-ready libraries in about 3 hours without the need for end-repair, adaptor ligations, shearing of the cDNA, or gel electrophoresis.
  • Uses random-primed cDNA synthesis to enable library preps from rRNA-depleted RNA or Poly(A)+ RNA from any animal, bacteria, or plant species.
  • Is barcode-capable for Illumina-compatible barcodes (available separately) or user-defined barcodes.
  • Produces mRNA-Seq libraries that demonstrate high concordance with MAQC qPCR data (Fig. 3).
  • Requires as little as 50 ng of rRNA-depleted RNA.
  • Generates highly mappable libraries from intact, partially degraded, and FFPE RNA samples (Table 2).

*Covered by issued and/or pending patents.


Figure 1

Table 1. ScriptSeq™ mRNA-Seq Library Preparation Kit (Illumina-compatible) generates libraries for directional sequencing in less than 3 hours. Times for each step are shown in hours:minutes.

Conventional mRNA-Seq Method ScriptSeq™ Method
Fragment RNA (1:00) Fragment RNA and synthesize di-tagged cDNA (1:40)*
Synthesize cDNA (4:30)
Ligate adaptors (2:00) Clean up cDNA (0:10)
Size-select from gel (1:30)  
Enrich library by PCR (1:00) Enrich library by PCR (1:00)
Total time: 10:00 Total time:
*single-tube reaction
Figure 1 (click to enlarge). Schematic overview of the ScriptSeq™ directional library preparation method. The process is complete in less than 3 hours, with no intermediate purification steps from RNA to di-tagged cDNA fragments.
Figure 2A
Figure 2B
Figure 2 (click to enlarge). Representative Bioanalyzer traces of input human brain RNA and corresponding ScriptSeq libraries. A) Poly(A)-enriched RNA; B) ScriptSeq library prepared from poly(A)-enriched RNA; C) rRNA-depleted RNA prepared using the Ribo-Zero Kit; D) ScriptSeq library prepared from Ribo-Zero rRNA-depleted RNA.
Figure 2C
Figure 2D

A. Intact UHRR and BrRR
Figure 3A
B. Fragmented UHRR and BrRR
Figure 3B
Figure 3. Correlation of gene expression between data obtained from EPICENTRE ScriptSeq™ libraries prepared and corresponding MAQC qPCR data. A) Libraries prepared from rRNA-depleted, intact UHRR and BrRR. B) Libraries prepared from partially fragmented, rRNA-depleted UHRR and BrRR. UHRR, Universal Human Reference RNA; BrRR, Brain Reference RNA.



Table 2. The ScriptSeq™ mRNA-Seq Kits generate high quality libraries from rRNA-depleted RNA, poly(A) RNA, and FFPE RNA samples. The specified RNAs were treated with EPICENTRE's Ribo-Zero™ rRNA Removal Kit (Human/Mouse/Rat) or a commercial poly(A) RNA enrichment kit. ScriptSeq (Illumina-compatible) libraries were prepared and sequenced on an Illumina® GAII sequencer. UHRR, Universal Human Reference RNA; BrRR, Brain Reference RNA.

RNA rRNA Removal Method Number of Reads Reads Mapped to Genome Uniquely Mapped Reads (%)
UHRR Ribo-Zero™ Kit 27,524,073 87.0% 21,929,999
UHRR Poly(A) enrichment 30,123,553 80.7% 21,845,097
BrRR Ribo-Zero™ Kit 31,998,097 88.0% 26,019,696
BrRR Poly(A) enrichment 17,763,706 79.7% 13,265,142
FFPE RNA (human breast tumor) Ribo-Zero™ Kit 26,960,546 85.6% 21,515,095


Catalog No. Size

ScriptSeq™ mRNA-Seq Library Preparation Kit (Illumina-compatible)
SS10906 6 Reactions
SS10924 24 Reactions

RNA-Seq Barcode Primers (Illumina-compatible)
RSBC10948 48 Reactions
Set of 12 Illumina-compatible barcodes.