The Ion Total RNA-Seq Kit v2 includes the reagents needed to prepare representative cDNA libraries for strand-specific RNA sequencing on the Ion GeneStudio S5, Ion Proton, or the Ion Personal Genome Machine (PGM) systems. The Ion GeneStudio S5 and Ion Proton systems are ideally suited for sequencing the whole transcriptome (ribosomal RNA depleted or polyA), while the Ion GeneStudio S5 and Ion PGM systems are suitable for viral and bacterial transcriptomes. Version 2 of the Ion Total RNA-Seq Kit is an improvement over the first generation kit. Magnetic bead-based purification replaces all of the filter cleanup steps and the total reaction time has been reduced to 6 hours.

Additional Features of the New Ion Total RNA-Seq Kit v2:

• Greater accuracy—SuperScript VILO and Platinum PCR SuperMix High Fidelity added for highest template fidelity
• Barcode compatible—works with Ion Xpress RNA-Seq Barcode 01-16 Kit for multiplexing
• Automation friendly—magnetic bead-based purification simplifies automation of library construction

As with the previous kit, the Ion Total RNA-Seq Kit v2:

• Preserves strand information—all mapped reads are aligned in the direction of transcription relative to the chromosomal strand
• Allows you to analyze different types of RNA—supports rRNA depleted total RNA, and poly(A) RNA

The Ion Total RNA-Seq Kit v2 is designed to make cDNA library preparation fast and flexible. It can be used to generate a representative cDNA library, flanked by specific sequences necessary for sequencing, from any type of RNA sample.

Designed as a complete solution with a common workflow, the Ion Total RNA-Seq Kit v2 combines optimized reagents and protocols for discovery of coding RNA, noncoding RNA, and alternative splice variants.

Whole transcriptome analysis
The whole transcriptome protocol enables construction of strand-specific libraries in approximately 5 hours. Starting with as little as 100 ng of total RNA, construct a library from 1 ng of poly(A) RNA or 25 ng of rRNA-depleted RNA following the RNA enrichment and library generation protocols provided in the manual. Because the libraries are not limited to cDNA derived only from poly(A) RNA, Ion Total RNA-Seq Kit libraries support a more thorough investigation of transcriptome complexity, capable of characterizing known and undocumented transcripts, including alternative splice variants, fusion transcripts, and SNPs.

Preserve strand information
Unlike methods that ligate adapters to double-stranded cDNA, the Ion Total RNA-Seq Kit v2 utilizes proprietary Ambion technology to attach the adapters in a directional manner that preserves strand information in the resulting libraries. In addition, both the 3' and 5' adapters are attached simultaneously, reducing ligation and clean-up steps.

Preserving strand orientation during library construction helps enable more accurate determination of the structure and expression level of transcripts, and can aid in the discovery of novel transcription regions from both the positive and negative genomic strands.

The Ion Total RNA-Seq Kit v2 is designed to create RNA libraries from up to 48 samples for whole transcriptome sequencing on the Ion GeneStudio S5, Ion Proton, or Ion PGM systems.