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Finkelstein Lab Wiki

Lab Freezer Database

Here is where you will find information on the lab database. This is run through the Filemaker Pro 12 application and updated as it goes by everyone in the lab.

Contents

Section titled “Contents”
  1. Plasmids
  2. Strains
  3. Racks (Proteins)

1. Plasmids

Section titled “1. Plasmids”

After making a plasmid, please fill in the following information:

  1. pIF number for the plasmid (This will be the next available pIF number. i.e. if the last plasmid put in was pIF20, yours is pIF21.)
  2. A description of the plasmid. Please include at least
    1. Name of the gene
    2. Original Vector
    3. Any primers used to make the new plasmid
    4. Orientation of the gene of interest
  3. Make a strain number following the guidelines below and give that number.
  4. Fill in your initials.
  5. List the resistance markers in the plasmid.
  6. Upload the sequence information to the database.
  7. The date made, which will be auto-filled by the program.

2. Strains

Section titled “2. Strains”

Storing e. coli strains:

  1. Mix 1 vol 80% glycerol (sterile) with 5 volumes of culture (overnight or exponentially growing).
  2. Snap freeze in LN2.
  3. Strains stored in this fashion should be viable >25 years later.

After making your plasmid and sequencing it, please save the strain that was used to make it and input the following data:

  1. IF number for the strain (This is the next available IF number)
  2. Strain type (DH5a, DH10bac, ArcticXpress, etc)
  3. The plasmid carried in the strain (please the pIF number and gene of interest)
  4. Resistance Marker
  5. Your initials
  6. The date made (which will be filled in automatically)

3. Racks (Proteins)

Section titled “3. Racks (Proteins)”

When a protein is purified or received externally, it should be aliquoted and stored in a box in the -80C freezer. The -80C freezer is labelled as being double-lettered(i.e. AA, BB, CC for each freezer that we get), and each rack in the freezer has an individual number. i.e. the first rack is 1, the second is 2, etc. Each rack contains 5 rows, which are numbered 1-5, and finally, each rack row has 4 box spaces, lablled a-d. So, the third row of the second rack in the first box would be labelled (2.3a).

So, find an empty space in the freezer and put your box of protein in there and then fill in the database with the following information:

  1. Rack location
  2. Date made
  3. Description of the protein in the box
  4. Your initials