Sequencing - 200G to >20g
HiSeq 2500
Genome Analyzer IIx
MiSeq
Sequencing + Arrays
HiScanSQ
Arrays - Low- to High-Plex
HiScan
iScan
BeadXpress
Real-Time PCR System
Eco Real-Time
VeraCode DASL Gene Expression Analysis Kits
| System: | BeadXpress Reader |
|---|---|
| Technology: | VeraCode |
| Assay: | VeraCode DASL Gene Expression Assay |
| Applications: | Custom Low- to Mid-plex Gene Expression Analysis |
| Species: | Any |
| Contents: | The kit includes DNA synthesis reagents, DASL assay reagents, a custom Oligo Pool for Gene Expression Assay (DAP), and VeraCode BeadPlates. |
The VeraCode cDNA-mediated Annealing, Selection, extension, and Ligation (DASL) Assay (VeraCode DASL Assay) combines the DASL Gene Expression Assay with the data quality of VeraCode technology. The DASL Assay is highly robust to degraded RNA and is ideal for profiling formalin-fixed paraffin-embedded (FFPE) samples.
The VeraCode platform leverages holographic bead technology to enable flexible multiplex assay deployment. VeraCode microbeads are glass cylinders with each bead type uniquely identified by an inscribed holographic code.
VeraCode beads with immobilized universal capture oligonucleotides are an ideal high-throughput readout platform for the DASL gene expression assay. DASL chemistry proceeds just as on the BeadArray platform, but VeraCode solution-based kinetics help reduce total assay time to 2 days.
The DASL Assay is similar to the Illumina GoldenGate Assay, with an additional reverse transcription step with biotinylated random primers to create activated cDNA.
Each transcript assay consists of three oligonucleotides: two assay specific oligos (ASO1 and ASO2) and one locus-specific oligo (LSO). After specific hybridization of these to the cDNA, enzyme-mediated extension provides further target specificity. Extended ASOs are ligated to the LSO, creating a template for universal PCR. The number of PCR templates created corresponds to the number of transcripts of a given type in the original sample. Universal PCR with dye-labeled primers creates a fluorescent product amenable to detection and quantification.
Each assay type is uniquely identified with an LSO-derived address sequence. Each address is complementary to capture oligos on the surface of one VeraCode bead type, inscribed with an identifying code. By incubating DASL products with VeraCode beads, specific assay types are identifiable during readout by their unique code.
- Only 100-200 ng of total starting RNA is required
- Gene profiling assays at 96-plex can be read at a rate of 80 samples per hour with high accuracy
Applications
Custom Low- to Mid-plex Gene Expression Analysis
For researchers interested in RNA analysis of specific genes or regions of interest, a customized solution can be developed with the DASL Assay. The robust DASL assay chemistry can generate high-quality data from samples with low RNA abundance or partial RNA degradation, such as formalin-fixed, paraffin embedded (FFPE) samples.
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