Fast Plasmid Slot Lysis and Gram-Negative Bacteria Ghost Preparation Protocol

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RecentlyAmara et al. (2013) introduced a simple protocol for evacuating microbial cells based on using the critical chemical concentration of effective chemical compounds as well as enzymes. Such protocol succeeded to evacuate the microbes from their cytoplasmic content. Amara introduced the idea of preparing the DNA and the protein from microbes using the same concept by using the minimum inhibition concentration of the used chemical compounds in the Sponge-like protocol. Slot lysis is another protocol applied for screening a large number of clones. In this study, a simple new protocol is introduced. The protocol guarantees fast and simple plasmid isolation from E. coli. Those who are interested in isolating plasmids from gram positive strains and full maintain the evacuated cells they can determine the MIC of SDS and NaOH in the lysis buffer.

The protocol issimple fast and use only the GET-solution and the SDS-NaOH solution of the alkaline lysis protocol. In 1 ml appendorf tube put 5 μl of the GET-solution and 10 μl of SDS-NaOH solution. During preparing the SDS-NaOH solution it is recommended to prepare it fresh and not put the SDS on NaOH directly but put them in sequence in the water. By the toothpick, touch a colony from the original plate till the colony stack well. Touch genteelly (did not allow full loss of the colony) the new LB medium plate and number the site of the touch with number. Transfer the toothpick which still contains the rest of the touched colony in an appendorf tube which contain both of GET-solution and the SDS-NaOH solution. Incubate for 10 minutes. One could keep the toothpick or allow slight vortex and remove it before the incubation period. Do that for as much as you could for the colonies in the original plates.