A) Schematic of assay for determining di-nucleotide preferences of Cas13a/b enzymes.
B) Collateral activity of LwaCas13a, CcaCas13b, LbaCas13a, and PsmCas13b on orthogonal di-nucleotide reporters.
C) Schematic of collateral activity of Cas12a activated by dsDNA.
D) Comparison of collateral activity and pre-amplification enhanced collateral activity (SHERLOCK) of LwaCas13a, PsmCas13b, and AsCas12a. The dotted line denotes 2e9 (aM), the limit of AsCas12a sensitivity without preamplification. Values represent mean +/− S.E.M.
E) Schematic of in-sample 4 channel multiplexing using orthogonal Cas13 and Cas12a enzymes.
F) In-sample multiplexed detection of ZIKV ssRNA, ssRNA 1, DENV ssRNA, and dsDNA 1 with LwaCas13a, PsmCas13b, CcaCas13b, and AsCas12a.
G) Schematic of in-sample multiplexed detection of S. aureus thermonuclease and P. aeruoginosa acyltransferase synthetic targets with LwaCas13a and PsmCas13b.
H) In-sample multiplexed RPA and collateral detection at decreasing concentrations of S. aureus thermonuclease and P. aeruoginosa acyltransferase synthetic targets with LwaCas13a and PsmCas13b.