Applications

• Generate directional RNA-Seq libraries, virtually free of rRNA, from as little as 100 ng of intact or partially degraded total bacterial RNA for Illumina® sequencing in one day.

The ScriptSeq™ Complete Kit (Bacteria) combines Epicentre's powerful Ribo-Zero™ technology and rapid ScriptSeq v2 RNA-Seq library prep procedure for optimized comprehensive preparation of RNA-Seq libraries in 1 day (Fig. 1). The ScriptSeq Complete Kit (Bacteria) requires 1-5 µg and the ScriptSeq™ Complete Kits (Bacteria) - Low Input utilizes 100 ng - 1 µg of total RNA.

The ScriptSeq Complete Kit (Bacteria) includes Ribo-Zero™ rRNA removal reagents for removal of 23S, 16S and 5S rRNA from Gram-positive and Gram-negative bacterial total RNA (Fig. 2), and ScriptSeq v2 RNA-Seq Kit for directional RNA-Seq.

Benefits

• One-day RNA-Seq library preparation procedure from Gram-negative and Gram-positive bacterial RNA samples.
• Produces directional (>97%) RNA-Seq libraries.
• Libraries typically contain <3% rRNA reads.
• Utilizes as little as 100 ng of intact or partially degraded total bacterial RNA.
• Enables paired-end, single-read and multiplex sequencing using Illumina® TruSeq™ Cluster kits and sequencing primers.
• Index/barcode capable. 48 ScriptSeq™ Index PCR Primers are available separately for preparing barcoded libraries.
• Applicable to a broad spectrum of Gram-positive and Gram-negative bacterial.

Figure 1. An overview of the ScriptSeq™ Complete Kit (Bacteria) process. Cluster-ready ScriptSeq RNA-Seq libraries, virtually rRNA-free, are made in 1 day from as little as 100 ng of intact or partially degraded total RNA.

Figure 2. Library composition of ScriptSeq™ Complete Kit (Bacteria) samples. ScriptSeq libraries were constructed from 5 µg of B. subtilis (A) or E. coli (B) total RNA samples and sequenced on an Illumina® GAIIx.