• 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

        "BEST" PCR—从质粒上扩增DNA的PCR条件

        互联网

        3545

        1、PCR reaction system:

        25 ng linear template (~6.5 kb)

        50 pmol each primer

        100 pmol each dNTP

        1X Promega Taq buffer (no Mg2+)

        1.5 mM MgCl2

        1 U Taq DNA polymerase in 50 ul final volume

        2、PCR programme:

        92°C / 2'

        92°C / 30"

        50°C or 55°C (depends on Tm of oligos) /30"

        72°C / about 2' per kb

        Go to 2, 15 times

        70°C/ 8'

        4°C,hold.

        Takes about 2 hours to complete.

        3、Notes:

        If you are using Pfu turbo, use its buffer (Mg already added) and decrease elongation to 1'/kb DNA. Note that this DNA is blunt ended and can be cloned directly (no purification necessary) into a phosphorylated vector. Taq made DNA needs to be AT vector cloned.

        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