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.