Product Data Sheet

Temperature sensitive replication (repA101ts) and conditional replicon (oriR101, requires Pir in trans for plasmid replciation); encodes lambda Red genes (exo, bet, gam); native terminator (tL3) after exo gene; arabinose-inducible promoter for expression (P_araB); encodes araC for repression of P_araB promoter; Ampicillin resistant.

Without antibiotic selection, this plasmid can be cured from a strain with growth at 37℃- 42℃. For plasmid maintenance, the strain should be grown at 30℃.

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.