TruSeq Total RNA Training
|Instrument||Recommended Number of Samples||Read Length|
|NextSeq 550 System||RNA Profiling: 6-20 samples per run (based on 20 million reads per
Transcriptome Analysis: 2-8 samples per run (based on 50 million reads per sample)
|2 x 75 bp|
|HiSeq 2500 System||RNA Profiling: 30-200 samples per run (dual flow cell; based on 20
million reads per sample)
Transcriptome Analysis: 12-80 samples per run (dual flow cell; based on 50 million reads per sample)
|2 x 75 bp|
|NovaSeq 6000 System||RNA Profiling (samples per run, dual flow cell): S1: 160, S2: 320, S4:
768. Limited by index combinations (dual). Based on 20 million reads.
Transcriptome Analysis (samples per run, dual flow cell): S1: 64, S2: 128, S4: 400. Limited by index combinations (dual). Based on 50 million reads.
|≤ 2 × 100 bp|
|TruSeq Stranded Total RNA||Illumina Stranded Total RNA Prep with Ribo-Zero Plus|
|Assay Time||11.5 hours||~7 hours|
|Automation Capability||Liquid Handling Robots||Liquid Handling Robots|
|Content Specifications||Captures coding RNA plus multiple forms of non-coding RNA||Captures coding RNA plus multiple forms of non-coding RNA|
|Description||For whole-transcriptome sequencing studies. The Total RNA version depletes cytoplasmic rRNA while the Total RNA Gold version depletes cytoplasmic and mitochondrial rRNA.||For whole-transcriptome sequencing studies of multiple sample types. Includes Ribo-Zero plus module for single-tube depletion of abundant transcripts from multiple species|
|Hands-On Time||5.5 hours||< 3 hours|
|Input Quantity||0.1 – 1 ug high-quality total RNA. Lower-quality samples may require further optimization.||1-1000 ng total RNA for standard-quality RNA samples. 10ng total RNA minimum recommended for optimal performance and FFPE samples|
|Mechanism of Action||Bead-based rRNA depletion, cDNA synthesis, and PCR||Enzymatic rRNA depletion, ligation-based addition of adapters and indexes|
|Method||Whole-Transcriptome Sequencing||Whole-Transcriptome Sequencing|
|Multiplexing||Low-throughput kits: Pool up to 12 samples. Or pool up to 24 samples with sets A and B together, High-throughput kits: Prepare 96 indexes samples using either combinatorial dual indexes or unique dual indexes (available soon)||Up to 384 Unique Dual Indexes (UDIs)|
|Specialized Sample Types||FFPE Tissue||FFPE Tissue, Low-Input Samples|
|Species Category||Human, Mouse, Rat||Bacteria, Human, Mouse, Rat, Virus|
|Species Details||rRNA Removal: Depletes human, mouse, and rat rRNA||Ribo-Zero Plus included for depletion of abundant transcripts from multiple species including: human cytoplasmic & mitochondria rRNA, mouse rRNA, rat rRNA, bacteria Gram +/- rRNA, human beta globin transcripts|
BaseSpace Sequence Hub helps the Biotechnology Resource Center Sequencing Core (BRC-Seq) at the University of British Columbia deliver high-quality sequencing data and additional value to its customers.Read More
Consistent, precise measurement of RNA abundance is reflected by high reproducibility between technical replicates. These technical replicates of FFPE tissue show high concordance, indicating robust library prep performance. Axes are log2(FPKM). R2 value is shown.
TruSeq Stranded Total RNA gives excellent coverage across the top 1000 expressed transcripts in both fresh-frozen (FF, top) and FFPE (bottom) tumor and matched normal breast tissue, with > 98% aligned stranded reads. X-axis: position along transcript, Y-axis = percent coverge of combined reads.
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