1. Clones (e.g. EST) are frequently stored in 96 well plates. If master
plates are low density bacterial cultures, incubate master plates sealed
over night at 37 °C. This step is not necessary if the master plates are high
density bacterial cultures.
2. To prepare replicate copies of the master plates, prepare sets of
96 well round bottom plates and add 100 ml of
LB containing antibiotics.
3. Spin the master plates briefly (1000 rpm for 2 minutes).
4. Partially fill a container with 100% ethanol. Dip the 96 pin replicating block in the alcohol. Remove from the ethanol bath and then flame the pins.
5. Allow the inoculation block to cool, then dip the replication tool in
the master plate, and then into the daughter plate. Repeat for each replicate
6. Cover the inoculated 96-well plates with lids and place into a "zip-lock"
bag containing a moistened paper towel and grow overnight at 37 °C. These will
be the working plates
7. To freeze working plates, add 50 ml 45% sterile glycerol
to each well and store at -80 °C.