Figure 2. crRNA screen. A. The IS711-crRNA-1-4 structure and
the corresponding target sequences. The target sites are highlighted in
blue, and PAM sequences are marked red. B. Product of Cas12a cleavage,
20 μl of each reaction was electrophoresed (2% agarose), M: DL 500 DNA
marker (Takara, Code No.3590Q), 1-4: crRNA-1-4, (RPA amplification
products were 340 bp, 261/79 for crRNA-1, 303/37 for crRNA-2, 310/30 for
crRNA-3, 174/166 for crRNA-4), N: negative control. Figure 2C Four
groups of crRNAs were screened by the quantitative fluorescence method.
The reaction was performed at 42oC for 40min, and the
signal was collected every minute. The reaction entered the plateau
phase at 8-10min. As shown in the figure, the endpoint fluorescence
value of crRNA-2 was slightly higher than that of crRNA-1 and crRNA-3,
significantly higher than crRNA-4.