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Figure S1. Non-specific DNase activity is conserved across Cas12 proteins. Figure S2. The RuvC domain is responsible for ssDNA trans-cleavage. Figure S3. Preference for Cas12b-mediated trans-activated cleavage of non-specific ssDNA. Figure S4. Sensitivity and specificity of AaCas12b-mediated DNA detection. Figure S5. Comparison of sensitivity of AaCas12b- and LbCas12a-based DNA detection. Figure S6. CDetection achieves sub-attomolar sensitivity in DNA detection. Figure S7. Develop CDetection platform. Figure S8. Accurate DNA detection using CDetection platform. Figure S9. Potential off-target analysis of CDetection. Figure S10. Rapid and accurate diagnostic applications of CDetection. Figure S11. Genetic variants from ClinVar that, in principle, can be detected by CDetection platform. (PDF 2218 kb)

Table S2. Sequences of targeting crRNAs used for in vitro RNA transcription. Table S3. Target sequences harboring various 5â ˛ PAM sequences used for in vitro DNA cleavage assay. Table S4. Protospacer sequences used for genome editing. Table S5. crRNA scaffold optimization and screening. Table S6. Primer sequences used for PCR amplification. Table S7. Frequency of Cas12a- and SpCas9-mediated targeted indel mutations at on-target sites in human 293FT cells. Table S8. Off-target analysis by targeted deep sequencing in the human 293FT cells. Table S9. Off-target analysis by whole genome sequencing (WGS) in human 293FT cells. (PDF 395 kb)

Supplementary Sequences. The humanized Cas12a coding sequences, protein sequences, U6-crRNA backbone sequences, mammalian and prokaryotic expression vector sequences used in this study. (PDF 183 kb)

Table S1. Nucleic acids used in this study. Table S2. The components of buffers tested for Cas12b-mediated DNA detection in this study. (PDF 293 kb)

Supplementary methods. (PDF 124 kb)

Review history. (DOCX 1323 kb)