1.3.4 Screening of the crRNA
The constructed Brucella  IS711 plasmid as the template, applying the RPA-Cas12a detection method mentioned in 1.3.3, and screening the four groups of crRNA (IS711-crRNA-1-4), respectively. The Cas12a cleavage products will be checked by electrophoresis on 2% agarose gel to observe the cleavage efficiency. Furthermore, observe the endpoint fluorescence value with a real-time PCR instrument. Select the most efficient crRNA by the above two approaches.