(r2) ProtocolsFreezingStrains < Lab

---+ 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. |
Barrick Lab > ProtocolList > ProtocolsFreezingStrains
Topic revision: r2 - 2011-06-02 - 05:00:24 - Main.AurkoDasgupta