Freezing Strains

Items

1. Obtain a bag 2mL cryovials, currently kept in the shelves above the colony counting machines.

2. Keep the bag closed with paperclips and in own workspace while not being used.

3. Obtain labels from bottom drawer of the cabinet below the message board.

4. Obtain cap circles from _.

Preparing Strains

1. Write the three letter initials, the strain number and the time on the label. Also include whatever other distinctive information about the strain that can be fit onto the label.

2. Stick label on tube, and cover it with clear tape for longevity. Also write the strain number on the cap.

3. Steps a and b can be done in either order, however, adding glycerol first is recommended.

a. Add 200uL of 80% glycerol (found amongst the stock solutions) each cryovial.

b. Add 1000uL of each revived strain to the appropriately labeled cryovial.

4. Label the box of cryovials and record the rack and freezer in which it is stored.

Updating the Database

1. Log in to barricklab.org, then log in to lab databases.

2. Click "strains" on the left, then "input." Fill in fields as shown in the chart. Note that the database uses minus signs (–), which it distinguishes from hyphens (-). On a mac, option+hyphen results in a minus sign. Simply put, Ara– is a valid entry, while Ara- is not.

column	example	description
strain	REL	Initials of strain designation
number	606	Number of strain designation
freezer	Barrick 1	Freezer where sample is located. Choose from list.
rack	Barrick 3	Rack where sample is located. Should be in format: [last name] [number]
box	Barrick 4	Rack where sample is located. Should be in format: [last name] [number]
container	2 ml glass vial	Type of container used to freeze sample. Choose from list.
well		Well number for samples frozen in microplates.
experiment	LTEE	Sample experiment. Choose from list.
population		Sample population in experiment.)
time	0	Sample time point in experiment.
M/C	C	Mixed population or Clone sample
clone		Designation of particular clone. Either a number or a letter.
description	Ara– ancestor clone used to begin the long-term experiment.	Full description of the strain. Include information about what strain it is derived from and whether this sample has been subcultured from different samples.
media	DM1000	Media strain was grown in for freezing.
cryoprotectant	Glycerol	Choose from list.
source	REL	Stock center or other lab strain is from.
frozen	2002-08-17	Date when sample was frozen.