Bifidobacterium animalis ( of 1 mM ice-cold sucrose-citrate buffe...)

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Bifidobacterium animalis

of 1 mM ice-cold sucrose-citrate buffer, dispense in tubes and incubate for 3.5 hours at 4 C.

Electroporation of cells:

Add 0.5-1.5 g plasmid DNA to 80 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into prechilled cuvette.
Wipe moisture from the cuvette and insert the cuvette into the device.
Electroporation:

Mode Prokaryotes O Voltage (V) 1,200 V Time constant (T) 5 ms
Dilute with 800 l outgrowth medium and incubate for 2.5 h at 37 C.
Plate onto selective MRS agar plates; incubate anaerobically for 2-3 days at 37 C.

Expected Results: Transformation efficiency up to 9.4 x 10⁴ transformants/g of DNA.

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