Ligo-miR EZ Assay Kit
Future Ligo-miR Tools
Ligo-miR EZ Assay Kit*
Ligo-miR EZ is an assay kit consisting of all probes, reagents, and buffers necessary to perform 30-plex microRNA profiling on 50 samples. Multiplex expression of 30 microRNA in each sample is accurately quantified using our innovative PCR-free, ligation assay.
The assay is performed in 3 simple steps. First, the capture ligation reagents are added to a total 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. With this simple method Ligo-miR EZ achieves sensitivity 100-1000x higher than microarray!
- 1,000X higher sensitivity than microarray (subattomole)
- Sensitivity rivals Nanostring and Illumina
High Sample Throughput
- To screen 96 samples with 30 microRNA read depth:
- Ligo-miR EZ - 1 well plate and 4 hours total time
- PCR - 30 well plates and 10 days total time2
- Microarray - 96 microarrays and 8 days total time3
- Nanostring - 8 cartridges and 3 days total time4
- 20X Reduction in Time Compared to Next Fastest Product
Figure 1. Ligo-miR EZ is able to achieve sensitivity down to 1e-18 moles with less than 20% exp-exp CV and 20% CV across microRNA using simple PAGE analysis.
- To perform 30 microRNA analysis:
- Ligo-miR EZ - Only 5 pipetting steps per sample
- PCR - 150 pipetting steps per sample
- Nanostring - Requires robotic preparation
- 30X Less Labor Compared to PCR-based Approaches
Figure 2. High specificity is achieved with no cross-talk between microRNA.
Lower Assay Costs1
- For each 30 microRNA test:
- Ligo-miR EZ - Only $15/sample
- PCR - 30 individual reactions costs $130/sample
- Microarray and Nanostring - Costs >$220/sample
- ~10-20X Cost Savings
Reduced Capital Investments
- Ligo-miR EZ can be performed using only a thermal cycler and gel apparatus
- Nanostring and Illumina platforms require exotic instrumentation (>$200k).
Figure 3. Raw PAGE image of 6-plex microRNA expression data. Each of the 4 lanes contains expression data for a single sample. Each band position represents a single microRNA with band intensity representing expression level.
1Specifications and prices from http://www.ambion.com/, http://www.invitrogen.com/, http://www.nature.com/app_notes/nmeth/2007/071601/full/an2256.html, http://www.nanostring.com/, 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