Does anybody know a short (i.e. < 12h) plug/Lysis protocol for Lactobacillus spp., especially L. crispatus? We have tried 30 min lysozyme (with/without prior cooking of cell Suspension for 20 sec), followed by plug casting and 3 hour incubation with proteinase K with very variable results. Addition of mutanolysin also did not improve the outcome. Resulting band patterns were sometimes fine for some L. crispatus strains but did not work out for others.

However, I still shy away from described long protocols like 16h lysozyme, overnight proteinase K and 16 h restriction.

Any shorter protocol would really be helpful.