Ligo-miR EZ Assay Kit

Advanced Ligo-miR Tools

Early Access Program

Ligo-miR EZ 19+5 Assay Kit*

The Ligo-miR EZ assay kit contains of all probes, enzymes, and buffers necessary to perform 24-plex microRNA profiling on 50 samples. Multiplex analysis of 24 microRNA is accurately performed in each reaction using our innovative PCR-free, ligation assay.

Ligo-miR EZ Assay Kit

Ligo-miR EZ 19+5 uses a semi-custom panel consisting of 19 fixed microRNA and 5 user selectable microRNA. The 19 fixed microRNA comprise 5 control microRNA (3 spike-in + 2 snoRNA) and 12 application specific microRNA (e.g. general cancer, breast cancer, cardiovascular, etc). The 5 variable microRNA are selected at the user's discretion to enable application specific tailoring. Users will then be sent a kit containing their semi-custom 19+5 panel. Fully custom panels can also be created.

Ligo-miR EZ Process

The assay is performed in 3 simple steps:

  • First, the capture ligation reagents are added to an RNA sample and incubated for 1 hour.
  • Second, the coding ligation reagents are further added to the mixture and thermal cycled  for 50 cycles.
  • Finally, the reaction mixture is analyzed by PAGE. Each lane represents a single sample where each microRNA is accurately identified by band position and expression level is precisely determined by band intensity.

High Sensitivity1

  • <1e-18 moles sensitivity using fluorescent PAGE detection (no PCR)
  • 50X linear amplification
  • Linear quantification across 4-5 orders of magnitude dynamic range
  • Absolute expression levels are easily obtained
  • Higher sensitivity than microarray
  • Sensitivity rivals Nanostring and Illumina

Fig 1. Linear quantification down to <1 amol.

Low Bias

  • High efficiency and low bias microRNA capture
    • 86% (AVG) ± 10% (SD) efficiency across across a representative 20 microRNA panel
    • No adapter pools necessary
    • Flex couple into other microRNA methods
  • Linear amplifcation = low amplification bias
  • Highly reproducible
    • Exp-Exp detection efficiency CV ~10-15%
  • High correlation to Taqman qPCR profiles

Ligo-miR EZ miRNA Capture

Fig 2. 20 microRNA panel captured at 86% (AVG)±10% (SD) efficiency.

High Specificity

  • No cross-talk across unrelated microRNA
  • Absolute discrimination of mature vs. pre-cursor microRNA
  • High specificity discrimination of closely-related microRNA families
    • Discriminates single nucleotide differences

Lig-miR EZ Let-7 family

Fig 3. High specificity discrimination of let-7 family.

High Sample Throughput

  • To screen 96 samples with 24 microRNA read depth:
    • Ligo-miR EZ - 1 well plate / 8 hours total time
    • PCR - 24 well plates / 8 days total time2
    • Microarray - 96 microarrays / 8 days total time3
    • Nanostring - 8 cartridges / 3 days total time4
  • 20X Reduction in Time Compared to Next Fastest Product

Ligo-miR EZ High Replicate Correlation

Fig 4. High reproducibility across replicate experiments.

Lower Assay Costs and Labor1

  • For each 24 microRNA test:
    • Ligo-miR EZ - Only $20/sample
    • PCR - 24 individual reactions costs $80/sample
    • Microarray and Nanostring - $250/sample
    • 24X less pipetting than qPCR
  • ~4-10X Cost Savings

Reduced Capital Investments

  • Ligo-miR EZ can be performed using only a thermal cycler and gel apparatus
  • Nanostring and Illumina platforms require proprietary instrumentation (>$200k).

Fig 5. PAGE image of 24-plex expression data from 4 cell lines. Each band represents a single microRNA with band intensity representing expression level.

*Patent Pending
1Specifications and prices from,,, Digital Multiplexed Gene Expression Analysis Using the Nanostring nCounter System in Current Protocols in Molecular Biology: John Wiley & Sons, Inc, and Direct Multiplexed Measurement of Gene Expression with Color-Coded Probe Pairs. Nature Biotechnology v26, p317-25.
2Assumes a single PCR machine, 4 hours per reaction, 3 reactions per day
3Assumes 12 hybridization chambers, overnight hybridization,
4Assumes cartridges run 24h a day