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

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

货号

SS10924

规格

24 Rxns

存储温度

-20℃

产品详情

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		 ~3:00
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.
A
Figure 2A		 B
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.
C
Figure 2C		 D
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
(79.7%)
UHRR Poly(A) enrichment 30,123,553 80.7% 21,845,097
(72.5%)
BrRR Ribo-Zero™ Kit 31,998,097 88.0% 26,019,696
(81.3%)
BrRR Poly(A) enrichment 17,763,706 79.7% 13,265,142
(74.7%)
FFPE RNA (human breast tumor) Ribo-Zero™ Kit 26,960,546 85.6% 21,515,095
(79.8%)

 

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.
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