SOLiD™ Total RNA-Seq Kit

Catalog number: 4445374

Applied Biosystems™  Related applications: RNA Sequencing | SOLiD® Next-Generation Sequencing

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The SOLiD® Total RNA-Seq Kit provides protocols and reagents for the construction of strand-specific libraries for small RNAs or the whole transcriptome. Specifically designed for use with the SOLiD® or 5500 Systems, the SOLiD® Total RNA-Seq Kit contains enough reagents to process up to 12 samples. In addition, both the small RNA protocol and the whole transcriptome protocol are compatible with the SOLiD® RNA Barcoding Kits. Features of the SOLiD® Total RNA-Seq Kit include:

• Paired-end reads enable accurate detection of splice variants and fusion transcripts
• Genomic DNA strand specificity is maintained, simplifying mapping to a reference database
• Uses total RNA, rRNA depleted RNA, or Poly(A+) RNA as input

Interrogate both small RNA and the whole transcriptome on the SOLiD® Sequencing System
For small RNA, libraries are generated from total RNA samples, or enriched for small RNA, for discovery via SOLiD® System sequencing. This is a robust method for whole genome discovery and the analysis of small non-coding RNA in general, and miRNA in particular. For whole transcriptome analysis, strand-specific libraries are constructed from total RNA, ribosomal RNA depleted total RNA or Poly(A+) RNA. Because the input RNA is not limited to Poly(A+) only, a more thorough investigation of the complexity of the transcriptome can be performed. For removal of ribosomal RNA, the RiboMinus™ Eukaryote System v2 or Low Input RiboMinus™ Eukaryote System v2 are recommended.

Genomic DNA strand specificity is maintained
High-throughput sequencing shows that non-coding RNA (ncRNA), typically representing a significant fraction of the eukaryotic genome, may be transcribed from both strands of the same genomic DNA region. The SOLiD® Total RNA-Seq Kit uses proprietary Ligase-Enhanced Genome Detection (LEGenD™) technology to convert RNA into a library of double-stranded cDNA molecules. Because the adapters are added simultaneously and in a directional manner, genomic DNA strand specificity is maintained. The final products are compatible with emulsion PCR on the SOLiD® System.
For Research Use Only. Not for use in diagnostic procedures.


For Use With (Equipment): SOLiD™
Library Type: cDNA Library
Workflow Step: Library Generation
Starting Material: RNA
Sequencing Type: Transcriptome Sequencing, Small RNA Sequencing
Product Size: 12 preps
Downstream Application: Next-Generation Sequencing

Contents & storage

Each SOLiD® Total RNA-Seq Kit includes:
• Nuclease-free water: 1.75 ml
• 10X RNase III Buffer: 20 µl
• RNase III: 20 µl
• SOLiD® Adaptor Mix: 30 µl
• Hybridization Solution: 40 µl
• 2X Ligation Buffer: 150 µl
• Ligation Enzyme Mix: 30 µl
• 10X RT Buffer: 50 µl
• dNTP Mix: 500 µl
• SOLiD® RT Primer: 30 µl
• ArrayScript™ Reverse Transcriptase: 20 µl
• 10X PCR Buffer: 660 µl
• AmpliTaq® DNA Polymerase: 110 µl
• SOLiD® 5′ PCR Primer: 100 µl
• SOLiD® 3′ PCR Primer: 100 µl
• WT Control RNA (1 µg/µl HeLa total RNA): 50 µl
• Small RNA Control (1 µg/µl human placenta total RNA): 10 µl
Store kit components at -20°C.


Manuals & protocols