丁香实验_LOGO
登录
搜索
    搜索
    提问
    我要登录
    ｜免费注册
    点赞
    收藏
    wx-share
    分享

    Purification of Inhibitor-Free Soil DNA Obtained by other Methods

    互联网

    621

    实验步骤

     

    1. Dissolve the DNA pellet with 200 ul of Elution Buffer or Buffer TE. Important: DNA pellet should be dissolved in Elution Buffer or Buffer TE(pH8.0).

    2. Add 50 ul inhibitor Removal Resin to the sample and vortex to mix well.

    Note: Completely resuspend HTR Reagent by shaking the bottle before use.

    3. Incubate at room temperature for 2 min and centrifuge at maxi speed ($13,000 x g) for 2 min to remove Inhibitor Removal Resin.

    4. Carefully transfer the supernatant into a new tube.

    Note: If supernatant still have dark color from soil, perform the HTR treatment again by repeating step 2-4.

    5. Transfer cleared supernatant to a new 2 ml tube.

    6. Add equal volume of XP2 Buffer, mix by vortexing. For example: if the sample from step 12 is 250 μl, add 250 μl Buffer XP2.

    7. Apply the sample from step 13 to a HiBind® DNA Column assembled in a 2 ml collection tube (supplied). Centrifuge at 10,000 x g for 1 min at room temperature. Discard flow-through liquid and re-use collection tube.

    8. Place the column into the same 2 ml collection tube from previous step and add 300 μl XP2 Buffer. Centrifuge at 10,000 x g for 1 minute. Discard the flow-through and collection tube.

    9. Place column into a new 2 ml collection tube (supplied) and wash by adding 700 μl SPW Wash Buffer diluted with absolute ethanol. Centrifuge at 10,000 x g for 1 minute. Discard flow-through liquid and re-use collection tube in next step.

    Note: SPW Wash Buffer is provided as a concentrate and must bediluted with absolute ethanol as indicated on the bottle and page 4.

    10. Repeat step 16 with a second 700 μl SPW Wash Buffer.

    11. Discard liquid and re-insert the column to the empty collection tube, centrifuge the column at full speed (13,000 x g) for 2 min at room temperature. This step is critical in removing traces of ethanol that will interfere with downstream applications.

    12. Place column into a clean 1.5 ml microcentrifuge tube (not supplied). Add 30-100 μl Elution Buffer directly onto the center of HiBind® matrix. Incubate at 65°C for 10-15 minutes.

    13. Centrifuge at full speed (13,000 x g) for 1 min to Elute DNA.

    14. Repeat elution step 12-13 with a second 30-100 μl Elution Buffer.

    ad image
    ad image

    相关产品推荐

    HB Western blotting Principles and Methods

    HB Western blotting Principles and Methods

    ￥216

    RNaseOff™ RNase Inhibitor，阿拉丁

    RNaseOff™ RNase Inhibitor，阿拉丁

    ￥447.90

    DNase I (RNase Free)，1000 U，阿拉丁

    DNase I (RNase Free)，1000 U，阿拉丁

    ￥537.90

    土壤DNA提取试剂盒（大提）-Soil DNA Isolation Plus Kit (100)

    土壤DNA提取试剂盒（大提）-Soil DNA Isolation Plus Kit (100)

    ￥33

    On-Site Instructor-Led Course: Biomek Introduction and Methods Programming (service)

    On-Site Instructor-Led Course: Biomek Introduction and Methods Programming (service)

    ￥93500

    相关问答

    问

    iTRAQ/TMT和Label-free技术都有哪些优缺点？

    3 回答 1488 围观
    2023-07-17

    问

    Label free 定量蛋白质

    3 回答 619 围观
    2023-03-29

    问

    Methods and Protocols系列丛书

    2 回答 366 围观
    2022-10-18

    相关方法

    DNA

    DNA印迹

    制备DNA

    推荐阅读

    Methods for microbial DNA extraction from soil for PCR amplification

    C1 Inhibitor: Quantification and Purification

    Purification of Endotoxin Free mAbs

    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