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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 that the plate contains 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.
  5. Mail DNA sample plates and cell sample plates separately. Please use procedures similar to that shown below:

    6.  

     

    Preparation of DNA microtiter plates Preparation of cell sample plates
    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 a letter "d" (lower case) to indicate that the plate contains DNA shown.
    3. After samples are added to plates, the DNA sample plate contents should be dried down.
    4. Seal tightly with Seal & Sample Aluminum Foil Lids (Beckman #538619) shown. A sealer tool shown may be used to ensure tight closure of the microtiter well contents.
    5. Label the sealing foil with your plate ID shown 
    6. Place the microtiter plate cover over the sealing tape/foil shown
    7. Tape down the microtiter plate lid shown.
    8. Ship on a Monday to allow delivery during normal hours of the work week (shipping DNA samples on dry ice is not necessary).
    9. Attach APHIS Courtesy Permit shown to exterior (highlight designated areas)
    10. Label shipment with:

      11.  

       
      • Contents have NO animal content, have NO bovine serum albumin, are NON-infectious and NON-hazardous. 
      • IN VITRO use (laboratory use only)
      • Indicate what the material is (recombinant DNA)
      • Identify the species from which the product is produced with the genetic inserts identified.
      • Identify the intended use of the product (to generate an extensive library of cereal grain gene sequences)
      • Country from which the product originates
      Ship to: 
      Carrie Rausch/Olin Anderson
      USDA, ARS, WRRC 
      800 Buchanan St.
      Albany, CA 94710-1105
      USA 

    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 a letter "c" (lower case) to indicate that the plate contains CELLS shown.
    3. Cell culture sample plates should have ~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. Place the microtiter plate cover over the sealing tape/foil shown
    8. Tape down the microtiter plate lid shown.
    9. "Flash freeze" cell culture by submerging plates in liquid nitrogen.
    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 areas)
    12. Label shipment with:

      13.  

       
      • Contents have NO animal content, have NO bovine serum albumin, are NON-infectious and NON-hazardous. 
      • IN VITRO use (laboratory use only)
      • Indicate what the material is (cells)
      • Identify the species from which the product is produced with the genetic inserts identified.
      • Identify the intended use of the product (to generate an extensive library of cereal grain gene sequences)
      • Country from which the product originates
      Ship to: 
      Carrie Rausch/Olin Anderson
      USDA, ARS, WRRC 
      800 Buchanan St.
      Albany, CA 94710-1105
      USA 

    For questions:
     
     

    about shipping and handling, contact:

           Carrie Rausch (crausch@pw.usda.gov, 510-559-5655).
    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".