Shreffler:Bacterial Cell Culture

Inoculation of Bacteria for Overnight Culture
Inoculation is the process of introducing a small amount of bacteria to growth medium in order to grow a large culture (e.g. to obtain more of the recombinant DNA that the cell has been transfected with).

Procedure
Note:Use a flame to keep the LB medium sterile
 * 1) Put 3 mL of LB medium into a 15 mL tube
 * 2) Add 1.5 μL carbinicillin (final concentration 50 μg/mL)
 * 3) With a sterile toothpick held by flamed tweezers, pick a colony off the plate and place the toothpick into the 3 mL of medium
 * 4) Place in shaker overnight (approximately 16 hours) at 37°C at a speed of 240 rpm
 * 5) Carefully remove and add 75 μL of carbinicillin to the two 150 mL flasks of LB medium previously prepared
 * 6) For the next overnight culture, add 300 μL of the first overnight culture to each of the flasks (1 to 500 dilution)
 * 7) Place in shaker overnight (approximately 16 hours) at 37°C at 240 rpm

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