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Getting males from a hermaphrodite population. This is a modified version of protocol originally written by Michael Koelle at Yale University.

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[Bio101] Making Males of C. elegans
[Bio101] 从秀丽线虫中分离雄性线虫

发育生物学 > 细胞生长和命运决定
作者: Fanglian He
4/20/2011, 12792 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.58

[Abstract] Getting males from a hermaphrodite population. This is a modified version of protocol originally written by Michael Koelle at Yale University.

[Abstract] 从雌雄同体的群落里得到雄性线虫. 这是一个修改版的步骤,最开始是由耶鲁大学的Michael Koelle 所写.

Materials and Reagents

  1. NGM plates

Equipment

  1. 25 and 30 °C incubators

Procedure

  1. Set up ~6 NGM plates (seeded with normal bacterial food E. coli OP50-1) with 10 L4 hermaphrodites each.
  2. Heat shock 4-6 h (no more than 6 h) at 30 °C.
  3. Move plates to 25 °C. You should get a few males per plate in the F1 generation.
    Note: To get more males, it is best to set them up with an excess of L4 hermaphrodites to ensure more males recover in the next generation.
  4. Male stocks should be maintained by selecting out an excess of males, along with a few L4 hermaphrodites in each generation (usually, male to hermaphrodite ratio is about 1: 10).

材料和试剂

 

1.        NGM 平板 (hyperlink)

 

设备

 

1.        25 and 30oC 培养箱

 

步骤

 

1.         ~6 NGM 平板 (seeded 用正常细菌需要的食物E. coli OP50-1) with 10 L4 每个雌雄同体.

2.        30 oC下热休克4-6小时(不超过6小时).  

3.        转移平板到25°C. 你应该在F1 代每个平板中得到少数雄性线虫.

注意: 想要得到更多雄性线虫, 最好把他们用一个过量的L4雌雄同体建立起来确保在下一代恢复更多的雄性线虫.

4.        雄性stocks 应该通过选择一个过量的雄性,与一些L4雌雄同体来维持, 每一个时代(通常,雄性与雌雄同体的比例是1:10.

 

 

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How to cite this protocol: He, F. (2011). Making Males of C. elegans. Bio-protocol Bio101: e58. DOI: 10.21769/BioProtoc.58; Full Text



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11/7/2012 8:25:19 AM  

I wasn't sure about this thank you!

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3/16/2012 10:24:22 PM  

Thanks for contributing. It's helped me undersantd the issues.

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