Current screening methods for recombinants include colony hybridization, PCR and plasmid preps. Each of these methods has disadvantages. Colony hybridizations require several days to a week and may involve the use of radioactivity. PCR reagents are expensive and the procedure requires lengthy set-up and reaction times.
Plasmid preps require many hours for cell growth and a laborious purification of mini-prep plasmids.
In addition, if the plasmid screening is done using commercially available plasmid prep kits, the cost of reagents rapidly becomes significant. As a result, DNA yields from bacterial lysis are much greater using Ready-Lyse Lysozyme, since DNA losses due to nonspecific binding are substantially less. Using this procedure, several hundred colonies per day can be screened for recombinants using only small amounts of reagents. The protocol for the in-well lysis procedure is shown in Table 1. Table 2 gives the composition of the protoplast buffer and lysis solution.