Growing Bacterial Cultures for DNA Plasmid Isolation

Growing Bacterial Cells

LB Broth
950ml water
bactotryptone 10g
5g yeast
10g NaCl

Alternatively, for LB broth, add 25g / L powder LB

when used as a medium for LB broth served in a thermos,
for thermos 2L, the maximum number of media to use is ~ 500ml, or from volume flask to enable the supply of oxygen for the bacteria.

Growing Bacterial Cells

Creating a permanent stock of bacterial cells
830uL of new-grown bacterial cultures
170uL of 85% glycerol
freeze in freezer 80 degrees

Growing cells from permanent stock for maxi-plasmid preparation
1. maintain the cells in the block freezer when taking them out
2. Bunsen burner flame with tweezers
3. Take a sterile pipette tips with a pair of tweezers to scoop cell
4. place in ~ 5 mL of broth containing the desired antibiotic
5. rocking overnight
6. take 500uL of culture and place in a 250 mL LB broth (flask 2L) rocked overnight
7. ready for the next day plasmid DNA isolation

Growing cells from the plate change

1. tweezers fire with a Bunsen burner.
2. The use tweezers to pick up a sterile pipette tips.
3. choose one colony.
4. dump the pipette tip into culture tubes with 2 mL of LB and shake overnight at 37 degrees C.