. Author manuscript; available in PMC: 2020 Apr 1.

Published in final edited form as: Nat Protoc. 2019 Sep 23;14(10):2986–3012. doi: 10.1038/s41596-019-0210-2

Table 1:

Comparison of two-step and one-pot formats of SHERLOCK

	Two-step SHERLOCK	One-pot SHERLOCK
Experimental time	30–60 minutes	15–30 minutes
Protocol optimization	Easy (each step can be individually optimized)	Difficult (more challenging compared to two-step)
Contamination risk	increased risk of surface contamination with pre-amplification reaction	decreased risk due to single tube reaction
Quantitative	Difficult (depends on finding time-point where separation is possible)	Easy (owing to real-time detection).
Sensitivity	zeptomolar range (single molecule per mL)	femto to attomolar range (1000–1 molecule(s) per μL). Variability in sensitivities have been observed which we largely attribute to the viscous RPA sample composition that can influence Cas13 detection.
Appropriate applications	End-point binary detection	time-sensitive applications (< 30 minutes)
	applications with challenging sample inputs (e.g., quick extractions from unpurified samples)	Quantitative applications high-throughput applications
		applications with increased contamination risk (repeated testing, large target amounts)