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Resources » Paper

Takao Ishidate et al. (2002) Worm Breeder's Gazette "A method of growing large synchronous population of C.elegans using chicken eggs."

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  • Comments on Takao Ishidate et al. (2002) Worm Breeder's Gazette "A method of growing large synchronous population of C.elegans using chicken eggs." (0)

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    Status:
    Publication type:
    Gazette_article
    WormBase ID:
    WBPaper00015806

    Takao Ishidate, & Craig Mello (2002). A method of growing large synchronous population of C.elegans using chicken eggs. Worm Breeder's Gazette, 17(3), 27b. Unpublished information; cite only with author permission.

    It is sometimes required to prepare a large synchronously growing population of worms for various biochemical or genetic assays. Use of chicken eggs to grow gram quantity of worms has been described (D.Baillie and R.Rosenbluth; WBG2(1)). We have found that by adding E.coli to blended chicken egg it is possible to grow a large population of worms synchronously on plates. In this method, freshly hatched L1 larvae are suspended in blended chicken egg solution supplemented with concentrated E.coli, and seeded onto standard NGM plates. Typically, 1 million worms can be grown synchronously on five, 150mm NGM plates. 1) Take one egg and break into 25ml of boiling water placed on magnetic stirrer. 2) Stir several minutes until yolk breaks. 3) Put the egg solution into Waring blender and blend for two minutes. 4) Transfer the homogenized egg solution to sterile tubes. (They can be frozen and stored at -80C.) 5) Mix egg solution and concentrated E.coli solution (OP-50 grown in Terrific Broth for one day is pelleted and then resuspended in equal volume of sterile M9). Usually we use the ratio of 1:9 = concentrated E.coli : egg solution. 6) Suspend 200,000 freshly hatched L1 larvae in 1~1.5ml of egg/E.coli mixture and inoculate onto 150mm NGM plate

    Affiliation:
    - Biotech II Suite217, Univ. of Massachusetts Med School, Worcester, MA 01605


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