• 我要登录|
  • 免费注册
    |
  • 我的丁香通
    • 企业机构:
    • 成为企业机构
    • 个人用户:
    • 个人中心
  • 移动端
    移动端
丁香通 logo丁香实验_LOGO
搜实验

    大家都在搜

      大家都在搜

        0 人通过求购买到了急需的产品
        免费发布求购
        发布求购
        点赞
        收藏
        wx-share
        分享

        Electro-Transformation of Yeast电转酵母【Upstate Medical University】

        互联网

        1777

        Amberg Lab ,Upstate Medical University

        1.Grow cells to 1X10E8 or OD600 of 1.2-1.3.

        2.Spin cells at 5,000rpm for 5 min, and wash pellets in an equal volume of ice cold water.

        3.Wash in 1/2 volume cold water.

        4.Wash in 1/25 volume ice-cold 1M. sorbitol.

        5.Treat cells with 25mM. DTT for 10 minutes at room temperature.

        6.Wash cells with 1M. cold sorbitol.

        7.Resuspend cells in 1/200 original volume cold 1M. sorbitol.

        8.Mix 50µl. of cell suspension with not more than 5µl. DNA (in low ionic strength buffer).

        9.Immediately tap cell/DNA suspension to the bottom of a 0.2cm cuvette, pulse at 1.5kV, 200Æ, 25 µFaradays (pulse time Å5 millisec).

        10.Immediately add 1ml. YEPD/1M. sorbitol and allow to recover at permissive temperature for one hour.

        11.Spin down cells and resuspend in 1ml. 1M. sorbitol.

        12.Plate on selective media.

        Note that 10ml. is the minimum culture volume and that cells can be stored for a few days if they are not DTT treated.

        ad image
        提问
        扫一扫
        丁香实验小程序二维码
        实验小助手
        丁香实验公众号二维码
        扫码领资料
        反馈
        TOP
        打开小程序