SHERLOCK optimization strategies. (A) Schematic of SHERLOCK workflow with areas of workflow optimization highlighted. (B) Comparison of direct cell lysis methods compared with column-based RNA purification from patient cells harboring PML::RARA short isoform at 30 minutes and 60 minutes of Cas13 detection. (C) Effect of decreasing RT-RPA time with fixed Cas13 detection time at 100 ng and 1 ng of input PML::RARA RNA. (D) Background-subtracted fluorescence intensity for detection of synthetic and cell line–derived BCR::ABL1 p210 isoforms using a 1-pot assay combining RT-RPA and Cas13a detection. (E) Demonstration of SHERLOCK assay readout in APL diagnostic sample using lateral flow strips.
Figure 6.

SHERLOCK optimization strategies. (A) Schematic of SHERLOCK workflow with areas of workflow optimization highlighted. (B) Comparison of direct cell lysis methods compared with column-based RNA purification from patient cells harboring PML::RARA short isoform at 30 minutes and 60 minutes of Cas13 detection. (C) Effect of decreasing RT-RPA time with fixed Cas13 detection time at 100 ng and 1 ng of input PML::RARA RNA. (D) Background-subtracted fluorescence intensity for detection of synthetic and cell line–derived BCR::ABL1 p210 isoforms using a 1-pot assay combining RT-RPA and Cas13a detection. (E) Demonstration of SHERLOCK assay readout in APL diagnostic sample using lateral flow strips.

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