Ambion® RNA-Seq Library Construction Kit for SOLiD® Next-Generation Sequencing
Ambion®, the leader in RNA analysis technologies, offers the first kit to streamline high-throughput sequencing of the whole transcriptome and small RNAs, while maintaining strand specific, on the Illumina® GAII platform. The Ambion® RNA-Seq Library Construction Kit compatible with the Illumina® GAII platform provides, in an optimized kit, all of the necessary reagents to discover coding RNAs, non-coding RNAs, and small RNAs and isoforms, as well as to show alternative splicing events. Based on the robust technology used in the SOLiD® Total RNA-Seq Kit, the Ambion® RNA-Seq Library Construction Kit compatible with the Illumina® GAII platform allows greater flexibility in RNA starting samples and improves sequencing results from the Illumina® GAII platform when compared to data from sequencing runs using the instrument manufacturer’s kits.
- Convenient—streamlined workflow interrogates both the whole transcriptome and small RNAs using one kit
- Versatile—use with total RNA, rRNA-depleted total RNA, mRNA, and small RNA
- Revealing—preserves strand specific relative to the chromosomal strand, aligning all mapped reads in the direction of transcription
Whole transcriptome and small RNA in one kit
With flexible input requirements and the ability to sequence a whole transcriptome or small RNA, the Ambion® RNA-Seq Library Construction Kit compatible with the Illumina® GAII platform is a convenient solution that appeals to the needs of any RNA-Seq experiment on the Illumina® GA II platform (Figure 1).
To perform whole transcriptome analysis, construct an RNA library compatible with paired-end sequencing in just 2 days, starting with as little as 100 ng of poly(A) RNA, 200 ng of rRNA-depleted RNA, or 200 ng of total RNA. Since each library is constructed to preserve information on chromosomal strand location, subsequent sequencing can detect known and unknown transcripts, including alternative splicing products, fusion transcripts, and SNPs (Figure 2).
A small RNA protocol enables the rapid construction of small RNA (microRNA ~10-40nt) libraries starting with total RNA or enriched small RNA. The Ambion® RNA-Seq Library Construction Kit compatible with the Illumina® GAII platform typically yields at least 50% mapping of all reads to miRBase (Figure 3).
Preserving Strand Specific Reveals New Data
Standard methods for constructing an RNA-Seq library fail to distinguish which strand a transcript is transcribed from. Consequently, sequencing data show equal levels of transcription from both strands. Without differentiation of areas of transcription between the two strands, valuable information is masked, such as the elucidation of antisense transcripts with regulatory function (which are different from those of the sense strand) and areas where alternative splicing or fusion transcript events occur between the two strands. In the example shown below (Figure 4), two distinct, adjacent, divergent transcripts are detected with strand-specific, genome-aligned reads. Without this strand information, the data would have appeared to represent a single transcriptional unit.
Based on the same proprietary technology used in the SOLiD® Total RNA-Seq Kit, the Ambion® RNA-Seq Library Construction Kit compatible with the Illumina® GAII platform most effectively constructs an RNA-Seq library that differentiates strands. Unlike other methods that ligate adaptors to double-stranded cDNA, the technology used in the Ambion® kit simultaneously attaches the 3’ and 5’ adaptors in a directional manner, reducing ligation and clean-up steps (Figure 5).
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- SOLiD® Total RNA-Seq Kit
- Illumina® TotalPrep™ RNA Amplification Kit
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.