E. coli K12 CJ236

Description

A suspension of E. coli strain CJ236 which has been grown in LB medium and brought to 50% glycerol.

Genotype

FΔ(HindIII)::cat (Tra⁺ Pil⁺ Cam^R)/ ung-1relA1 dut-1 thi-1 spoT1 mcrA

Properties and Usage

Antibiotic Resistance

·cam

Growth Media

LB

Growth Temperature

37 °C

Growth Medium and Conditions

1. Streak out the strain on LBagar containing chloramphenicol (15 µg/ml) to ensure that you start withan F⁺ host.
2. Do not include chloramphenicol in liquid media: the strain does notgrow well.
3. CJ236 is non-suppressing. Do not use M13 vectors that carry amber mutations(mp8, 9 and some mp10 and 11).
4. Store plates at 4°C in the dark.

Notes

CJ236 is dut^-ung^- andcontains an F-factor carrying a selectable marker (chloramphenicol resistance).It was made by taking an isolate of BW313 that had spontaneously become F^- andintroducing pCJ105, the F´ Cm^R construct (1,2). For use inpreparing uracil-containing M13 templates (3,4).

References

1.Joyce, C. and Grindley, N. (1984). J. Bacteriol.. 158,636-643.

2.Raleigh, E.A., Lech, K. and Brent, R. (1989). F.M. Ausubel et al.(Ed.), CurrentProtocols in Molecular Biology. x. New York, Unit 1.4: PublishingAssociates and Wiley Interscience.

3.Kunkel, T.A., Bebenek, K. and McClary, J. (1991). J. Miller(Ed.), MethodsEnzymol.. 204, 125-139. San Diego: Academic Press.

4.Kunkel, T.A. et al. (1987). R. Wu and L. Grossman(Ed.), MethodsEnzymol.. 154, 367-382. San Diego: Academic Press.

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