• 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

        FFPE Direct PCR Kit

        互联网

        931

        实验步骤

         

        1. Protocol for FFPE Extraction

           1) cut a approximately 10 mg FFPE sample into a 2 ml collection tube or suitable vessel, cut off the Samples tiny and remove the paraffin wax as far as possible.

           2) approximately 10 mg FFPE sample add 95 ul FP1 Buffer and 5 μl FP2 to the collection tube. Close the tube and vortex briefly. Make sure the sample is covered by the Extraction Solution.

           3) Incubate at 60°C for 30-60 minutes.

           4) Incubate at 90°C for 60 minutes.

           5) Add 100 μl FP3 Buffer and vortex to mix.

           6) Store the extraction at 2-8°C.

        2. PCR Protocol

        This protocol serves as a guideline for PCR amplification. Optimal reaction conditions, such as incubation times, temperatures, and amount of template DNA, may vary and must be individually determined.

           1) Thaw primer solutions. Keep on ice after complete thawing, and mix well before use.

           2) Mix the PCR Master Mix by vortexing briefly. It is important to mix the PCR Master Mix before use to avoid localized differences in salt concentration.

           3) Prepare one of the following reaction mixes on ice: (For a 25 μl reaction volume)

           4) Gently mix the reaction and spin down in microcentrifuge.

           5) Set up program for a routine PCR reactions:

              Initial Denaturation 94-95°C for 1-5 min

              25-40 cycles 94-95°C for 30 sec

                                     45-70°C for 10-30 sec

                                     72°C for X min(1min/kb)

               Final extension 72°C for 7 min

               Final soak 4-10°C

           6) For a simplified hot start, proceed as described in step 7. Otherwise, place the PCR tubes in the thermal cycler and start the cycling program.

           7) Simplified hot start: Start the PCR program. Once the thermal cycler has reached 94°C, place the PCR tubes in the thermal cycler. In many cases, this simplified hot start improves the specificity of the PCR.

        ad image
        ad image

        相关产品推荐

        伯优品牌|FFPE RNA提取试剂盒(离心柱法)

        伯优品牌|FFPE RNA提取试剂盒(离心柱法)

        ￥3520

        SuperRT One Step RT-PCR Kit，阿拉丁

        SuperRT One Step RT-PCR Kit，阿拉丁

        ￥1797.90

        伯优品牌|FFPE RNA提取试剂盒(自动化仪)

        伯优品牌|FFPE RNA提取试剂盒(自动化仪)

        ￥3520

        FFPE 样本 | 空间转录组测序

        FFPE 样本 | 空间转录组测序

        ￥29999

        FFPE样本｜细胞核提取试剂

        FFPE样本｜细胞核提取试剂

        ￥3535

        相关问答

        问

        pierce albumin depletion kit

        157 围观
        2024-09-14

        问

        请问 5 min TA/Blunt-Zero Cloning Kit 产品中的Blunt载体连接3Kb的片段，为什么连接不上?

        2 回答 1149 围观
        2023-05-20

        问

        石蜡样本（FFPE）是否可以进行蛋白质组学？

        4 回答 1358 围观
        2023-02-20

        相关方法

        解锁 Visium FFPE 实验全流程

        时空转录组 FFPE 样本收样标准

        使用 HC ExpressArray Kit 进行杂交与检测实验

        推荐阅读

        Plant Seed Direct PCR Kit

        Dynabeads® mRNA DIRECT Kit

        Dynabeads® mRNA DIRECT Micro Kit - mRNA isolation for RT-PCR amplification

        无忧采购 轻松科研

        无忧采购 轻松科研

        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