• 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

        Restriction Enzyme Digestion of DNA

        互联网

        1915

        Materials:

        10X restriction enzyme buffer (see manufacturer's recommendation)

        DNA

        sterile water

        restriction enzyme

        phenol:chloroform (1:1)

        1.Add the following to a microfuge tube:

        2 μl of appropriate 10X restriction enzyme buffer

        0.1 to 5 μg DNA

        sterile water to a final volume of 19 μl (Note: These volumes are for analytical digests only. Larger volumes may be necessary for preparative digests or for chromosomal DNA digests.

        2.Add 1 to 2 μl (3 to 20 units) enzyme and mix gently. Spin for a few seconds in microfuge.

        3.Incubate at the appropriate temperature (usually 37E C) for 1 to 2 hours.

        4.Run a small aliquot on a gel to check for digestion.

        5.If the DNA is to be used for another manipulation, heat inactivate the enzyme (if it is heat labile) at 70 E C for 15 min, phenol/chloroform extract and ethanol precipitate, or purify on Qiagen DNA purification column.

        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