Fig. 2: Universal cRASL-seq assay for pathogen-associated RNA analysis.

Each reference organism was serially diluted into PBS and added directly to the lysis buffer and probe pool. NLC, No Ligase Control; NTC, No Template Control. The extraction free protocol of Fig. 1C was performed with all 80 probe sets in a single pool. Molecular Equivalents are calculated by normalizing read counts to a PCR spike-in sequence of known copy number. Detection of aggregate probes' read counts at a signal >10× the NTC was used to calculate the assay’s limit of detection for each organism.