The invention relates to the field of
bacterial colony screening, in particular to a bacterium screening
reagent, a
screening method, a kit and application. The invention provides a
bacteria dissolving
reagent, which is prepared from the following components: 10 to 30 mM of
Tris-HCl, 10 to 20 mM of EDTA (
Ethylene Diamine Tetraacetic Acid) and 0.01 to 0.55 mg / mL of
lysozyme. According to the invention, the
CRISPR Cas12a technology is applied to
bacterial colony screening for the first time, and is improved and optimized based on the technical principle, so that the
CRISPR Cas12a technology is more suitable for high-
throughput screening. In
plasmid design, a PAM sequence is introduced to the
tail end of a
gene, so that all crRNA sequences can simultaneously recognize a
gene segment and a vector segment. In addition, automatic equipment and a large-batch screening
reaction system and
reaction conditions are utilized, so that the method is fully suitable for an automatic implementation scheme of
bacterial colony screening, the screening time is greatly shortened, and the strain breeding speed and the strain screening efficiency are greatly improved.