HTG EdgeSeq assays employ nuclease protection to measure RNA. Target-specific protection probes with universal priming sites (wings) hybridize to target transcripts and wingman (1). S1 nuclease is added to digest non-hybridized probes and RNA, resulting in a 1-to-1 ratios of probes and target RNAs (2).
After S1 nucleuse inactivation (3), the remaining probes are amplified with primers carrying sequencing adaptors and molecular barcodes (4). The resulting PCR products are purified, quantitated, and combined to make a sequencing ready library (5). HTG EdgeSeq assays do not require processing steps such as reverse transcription, adenylation, or adaptor ligation.
Steps 1 - 3 are fully automated on the HTG EdgeSeq system.
Our chemistry supports a wide variety of sample types, including formalin-fixed, paraffin-embedded (FFPE) samples.
Our multiplexed gene expression assays leverage the power and sensitivity of next-generation sequencing (NGS). Measure the expression levels of a few or thousands of genes with minimal sample input. HTG EdgeSeq assays generate results from clinically relevant samples, such as FFPE tissue, to support both research and diagnostic objectives.
Learn more about the HTG EdgeSeq Chemistry.
Page last updated March 11, 2020