• 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

        PCR Amplification of DNA

        互联网

        1576

        Materials:

        sterile water

        10X amplification buffer with 15mM MgCl2

        -10 mM dNTP

        50 μM oligonucleotide primer 1

        50 μM oligonucleotide primer 2

        5 unit/μl Taq Polymerase

        template DNA (1 μg genomic DNA, 0.1-1 ng plasmid DNA) in 10 μl

        mineral oil (for thermocyclers without a heated lid

        1. Combine the following for each reaction (on ice) in a 0.2 or 0.5 ml tube

        10X PCR buffer 10 μl

        Primer 1 1 μl

        Primer 2  1 μl

        dNTP 2 μl

        template DNA and water 85.5 μl

        Taq Polymerase 0.5 μl

        2. Prepare a control reaction with no template DNA and an additional 10 μl of sterile water.

        3. If the thermocycler does not have a heated lid, add 70-100 μl mineral oil (or 2 drops of silicone oil) to each reaction.

        4. Place tubes in a thermal cycler preheated to 94°C.

        5. Run the following program:

        94°C 1 min

        55°C 1 min or annealing temperature appropriate for particular primer pair

        72°C 1 min (if product is <500 bp), 3 min (if product is >500 bp)

        for 30 cycles.

        Program a final extension at 72°C for 7 min.

        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