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International Triticeae EST Cooperative (ITEC)

DNA & Clone Repository

Please use the following steps to submit your DNA and Clones (see images where available shown):
  1. For repository purposes, clones and DNA should be submitted in 96 well microtiter plates (Columns 1-12, Rows A-H). Preferred plates are Nunc Catalog No. 163320 shown, but any standard microtiter plates are acceptable.
  2. Plates should be labelled by the 3-letter lab identifier (see data submitting page), a 3-digit consecutive number of the plates from each lab, and "c" or "d" (lower case) to indicate the plate containing cells or DNA:


    3. For example, whe is the lab code for Olin Anderson's laboratory - thus, the 13th plate pair from Olin's lab will be labelled WHE013d (for DNA) and WHE013c (for cells). Labelling the plates with upper case letters will help prevent mistaking things like "l" for "i", etc... Other EST projects have reported 15-20% error rates in keeping track of clones. It will be important to be careful/fussy about labelling and tracking.

  3. The designation of individual sequences will consist of the lab code, the plate number, a period, the position on the plate of the clone/DNA (i.e., column and row), whether it is a forward or reverse sequencing reaction (F or R), and the date (format YYMMDD) that the sequence was generated. Thus, the plate information will be integrated into the sequence names, so labelling is important.


    4. For example, WHE013.D07F990405 is the sequence from the clone/DNA in position D-07 on plate #013 from Olin Anderson's laboratory, sequenced on April 5, 1999 using forward primers.

  4. Plates must be sealed to prevent cross-contamination during shipment with Seal&Sample aluminum foil sealing tape. Please use procedures similar to that shown below:

    5.  

     

    1. Use 96-well microtiter plates; preferred plates are Nunc Catalog No. 163320 shown.
    2. Be sure microtiter plates are labelled by the 3-letter lab identifier, a 3-digit consecutive number of the plates from each lab, and "c" or "d" (lower case) to indicate the plate containing cells or DNA shown.
    3. DNA sample plate contents should be dried down. Cell culture sample plates should have at least 100 µl culture volume with glycerol added to 15% (v/v) concentration.
    4. After samples are added to plates, seal tightly with Seal & Sample Aluminum Foil Lids (Beckman #538619) shown.
    5. A sealer tool shown may be used to ensure tight closure of the microtiter well contents. Liquid should not be able to leak out of any of the wells; it is advised that you test your sealing procedure first using water with dye.
    6. Label the sealing foil with your plate ID shown
    7. "Flash freeze" your cell culture plates in liquid nitrogen.
    8. Place the microtiter plate cover over the sealing tape/foil shown
    9. Tape down the microtiter plate lid shown.
    10. Immediately place shipment on dry ice (enough for 4- to 5-days) and ship on a Monday to allow delivery during normal hours of the work week.
    11. Attach APHIS Courtesy Permit shown to exterior (highlight designated area)
    12. Label shipment with:

      13.  

       

       
      Contents are NON-infectious, NON-hazardous, have NO animal content, and have NO bovine serum albumin.
       
       
      Ship to: 
      Carrie Rausch/Olin Anderson 
      USDA, ARS, WRRC 
      800 Buchanan Street 
      Albany, CA 94710-1105 
      USA


       
For questions:
 
 
about shipping and handling, contact:

       Carrie Rausch (crausch@pw.usda.gov, 510-559-5785).
about computer set-ups and sequence handling, contact:

       Gerard Lazo (lazo@pw.usda.gov, 510-559-5640).
other matters, contact:

       Olin Anderson (oandersn@pw.usda.gov)
 
 
Note: Numbering such as "001" and "002" are used in these instances to assist alignments and sorting of data by computer. This is currently in conflict with the "Catalogue of Gene Symbols for Wheat".