• 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

        Yeast Genomic DNA Isolation

        互联网

        780

         

        Yeast Genomic DNA Isolation

        David Amberg

        1. Grow 100 ml culture to about 1x10 to the 8th
        2. Spin 4 x 15 mls of culture down about 3k x 3 min.
        3. Resuspend entire pool in about 6 mls ddH2O, aliqoute into 1.5ml microfuge tubes and spin down.
        4. Aspirate off super and vortex pellet till suspended.
        5. Add 200ul soln A, add 200ul Phenol/CHCl3 and 0.3g acid washed glass beads.
        6. Vortex 2 min, add 200ul TEpH8 and spin at max speed for 5 min.
        7. Transfer super to a new tube and add 1ml 100% EtOH, mix by inversion.
        8. Spin 2 min, pour off super and recon in 0.4ml TEpH8 + 30ug/ml RNaseA.
        9. Incubate 5 min x 37¡C, add 18ul 5M NH4OAc and 1ml 100% EtOH and store at -20¡C for a few hrs.
        10. Spin down DNA 10 min and recon each pellet in 25-50ul or whatever convenient volume.
        • Solution A:
        • 200ul neat or 2ml 10% Triton X-100 (2%)
        • 1ml 10% SDS (1%)
        • 200ul 5M NaCl (0.1M)
        • 100ul 1M Tris pH8 (0.01M)
        • 20 ul 0.5M EDTA (1mM)
        • Water to 10ml

         

        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