Library Preps - RNA

Illumina Stranded Total RNA Prep, Ligation w/ Ribo-Zero Plus or Ribo-Zero Plus Microbiome

  • Detection of coding & non-coding regions
    • Global expression profiling
    • Novel feature detection
    • ncRNA discovery & characterization
    • Alternative splicing detection
    • Biomarker identification
  • Suitable for:
    • High-quality samples
    • Degraded samples
    • FFPE samples
  • Required total RNA input (in a volume ≤ 10 μL):
    • 1 - 1000 ng high-quality purified total RNA (RIN ≥ 9)
    • 10 - 100 ng low-quality RNA (RIN ≥ 2) or FFPE (DV200 > 55%) samples
    • 10 ng minimum for optimal quality & FFPE samples
    • RNA samples must be treated with DNase to minimize the contribution of sequence reads derived from residual genomic DNA in the sample
  • Includes:
    • Ribo-Zero Plus for multi-species rRNA depletion
      • Probes for Human Cytoplasmic rRNAs (28S, 18S, 5.8S, 5S), Human Mitochondrial rRNAs (12S, 16S), Human Beta Globin transcripts (HBA1, HBA2, HBB, HBG1, HBG2), Mouse and Rat rRNA (16S, 28S), Gram(-) Bacterial rRNAs (E. coli: 5S, 16S, 23S), Gram(+) Bacterial rRNAs (B. subtilis: 5S, 16S, 23S)
      • Custom probes can be spiked in (customer purchases separately)
        • Great success with custom probes for C. elegans, Pseudomonas aeruginosa, Streptococcus pyogenes
    • cDNA synthesis
    • 10 bp unique dual indexes (Illumina RNA UD Indexes Sets, Ligation)
      • Multiplexing of up to 384 samples

Illumina Stranded mRNA Prep, Ligation

  • Detection of the coding transcriptome w/ Poly-A tails
    • Differential expression profiling
    • Isoform identification
    • Allele-specific expression
    • Fusion detection & discovery
    • Detection of low expressed mRNA
  • Best suited for:
    • High-quality samples
    • Not compatible with FFPE samples
  • Required total RNA input (in a volume ≤ 25 μL):
    • 25 - 1000 ng high-quality purified total RNA (RIN ≥ 9)
  • Includes:
    • Poly-dT based mRNA capture
    • cDNA synthesis
    • 10 bp unique dual indexes (Illumina RNA UD Indexes Sets, Ligation)
      • Multiplexing of up to 384 samples

Other Library Preparation Options

  • Requests for projects requiring significant modifications to existing protocols or library preparation options not already listed here will be discussed with the customer on a per-project basis.

Notes & Resources