Product Data Sheet

Description

E. coli SM10 λpir strain

E. coli SM10 λpir strain contains the pir gene (lysogenized with λpir phage) and it is designed for cloning and propagation of plasmids with R6K origin of replication sequence (such as pUT mini-Tn5 vectors). The plasmids containing the R6K origin of replication require the R6K-spicific replication protein lpir gene for propagation. The genotype of the strain is KmR, thi-1, thr, leu, tonA, lacY, supE, recA::RP4-2-Tc::Mu, pir.

Genotype

The genotype of the strain is KmR, thi-1, thr, leu, tonA, lacY, supE, recA::RP4-2-Tc::Mu, pir.

Recovery

1.    Obtain an LB agar plate with the appropriate antibiotic.

2.    Using a sterile pipette tip, touch the bacteria growing within the punctured area of the stab culture. (A sterilized wire loop or sterile toothpick can be used in place of a sterile pipette tip.)

3.    Run this tip lightly over a section of the plate, as shown in the figure, to create streak #1.

4.    Using another sterile pipette tip, pass through streak #1 and spread the bacteria over a second section of the plate, to create streak #2.

5.    Using a third sterile pipette tip, pass through streak #2 and spread the bacteria over the last section of the plate, to create streak #3.

6.    Grow overnight in a 37 ^o C incubator (unless a different growth temperature is indicated on the plasmid datasheet).

7.    In the morning, single colonies should be visible. If the bacterial growth is too dense, re-streak onto a new agar plate to obtain single colonies.