JM101菌株/感受态细胞-BioVector NTCC保藏中心

大肠杆菌 JM101菌株/感受态细胞

Description:

 Asuspension of E. coli K12 JM101 which has been grown in minimal media and broughtto 50% glycerol.

Genotype:

                     F´traD36 proA+B+lacIqΔ(lacZ)M15/Δ(lac-proAB) glnV thi

                     Notes:

                     Storage at–70°C is recommended for periods longer than 30 days. Avoid repeated freeze/thawcycles.

                     References:

                     1. Messing, J. (1979) A multipurpose cloningsystem based on the single-stranded DNA bacteriophage M13, in Recombinant DNA TechnicalBulletin (NIH) Volume 3, Number 2.

                     2. Yanish-Perron et al. (1985) Gene33,103–119.

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.

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