• 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

        MINI PREP BY BOILING

        互联网

        927

        Pellet 1.5 ml of an overnight culture at 12,000 rpm in Eppendorf centrifuge at RT for 3 min.

        Resuspend bacterial pellet in 350 µl of STET buffer.

        Add 25 µl of freshly prepared solution of lysozyme (10 mg/ml in 10 mM Tris-Cl, pH 8).

        Mix by vortexing for 3 sec.

        Place the tube in a boiling water bath for exactly 40 sec.

        Centrifuge the lysate at 12,000 g for 10 min at room temp.

        Remove the glob of bacterial debris with a sterile toothpick.

        Add to the supernatant 40 µl of 2.5 M NaOAc (pH 5.2) and 420 µl of isopropanol.

         Mix by vortexing and store the tube for 5 min at room temp.

        Centrifuge at 12,000 g for 5 min at 4℃.

        Remove the supernatant and wash with 1 ml 70% ethanol.

        Dry the pellet and resuspend in TE (pH 8) with RNase A (20 µg/ml).

        STET buffer:

        0.1 M NaCl

        10 mM Tris-Cl, pH 8.0

        1 mM EDTA, pH 8.0

        5% Triton X-100

        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