2.5 Evaluation of the sensitivity and fluorescence intensity of
CRISPR Cas12a Detection
CRISPR Cas12a detection assays contained 50nM LbCas12a, 100 nM crRNA,
0.5 μl RNase inhibitor, 2 μl NEBuffer 2.1, FAM-BHQ1-labeled ssDNA
reporter and various amount of dsDNA template p3xFLAG-CMV-7.1-p72. The
sample without dsDNA plasmid template was set as negative control.
Reactions were incubated at a constant temperature of
37oC for 30 min and the fluorescent intensity was
measured by ABI QuantStudio 5. In addition, fluorescence was observed
with a UV transilluminator.