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

    E.Z.N.A.® Dried Blood DNA Spin Protocol

    互联网

    907

    实验步骤

     

    1. Cut or punch out the blood spot from the filter paper (up to 200ul of blood can be used per spot). Tear or cut the filter paper into small pieces and place them into a microfuge tube.

    2. Add 250ul of PBS Buffer, and incubate at 65℃ for 1 hour, while vortexing every 20 min for proper mixing.

    3. Add 25μl of OB Protease stock solution and mix well. Incubate for 30 min at 65℃ with occasional mixing.

    Note: Step 6 can be performed during incubation time.

    4. Centrifuge at 13,000 x g for 5 min at room temperature.

    5. Transfer the supernatant to a clean microfuge tube and add ONE volume of Buffer BL followed by ONE volume of absolute ethanol. Vortex thoroughly to mix. Collect any drops remaining on the lid by briefly centrifuging.

    6. Insert a HiBind® DNA Mini Column into a 2 ml collection tube (provided). Add 100ul Equilibration Buffer into the column. Let the column sit for 4 minutes at room temperature. Spin at maximum speed for 20 seconds.

    7. Transfer the lysate from step 5 into the column, and centrifuge at 10,000 x g for 1 min to bind the DNA. Discard the flow-through liquid and the collection tube.

    8. Place the HiBind® DNA Mini Column into a NEW 2 ml collection tube. Add 500:l of Buffer HB to the column, and centrifuge as above. Discard the flow-through and reuse the collection tube in the following step.

    9. Place the HiBind® DNA Mini Column into the SAME 2 ml collection tube from step 7, and wash by pipetting in 700 μl of DNA Wash Buffer diluted with ethanol. Centrifuge as above. Dispose of the collection tube and flow-through liquid.

    NOTE: DNA Wash Buffer is provided as a concentrate and must be diluted with absolute ethanol as indicated on the bottle, and on page 4. If refrigerated, the diluted DNA Wash Buffer must be brought to room temperature before use.

    10 Using a NEW collection tube (provided), wash the column with a second 700 μl of DNA Wash Buffer diluted with ethanol. Centrifuge as above. Discard flow-through liquid, and reuse the collection tube in the following step.

    11. Place the empty column into the same 2 ml collection tube from step 10. Centrifuge at maximum speed (13,000 x g) for 2 minutes to completely dry the column. Discard the flow-through and the collection tube.

    NOTE: This step is crucial for ensuring optimal elution in the following steps.

    12 Place the column into a sterile 1.5 ml microcentrifuge tube, and add 100-200ul of preheated Elution Buffer (10mM Tris-HCl, pH 8.5, 65℃). Allow tubes to sit for 5 min at room temperature.

    13. To elute DNA from the column, centrifuge at maximum speed (15,000 x g) for 1 min. Retain flow through containing the DNA. Discard column, and store the Eluted DNA at -20℃.

    NOTE : Blood Spots from finger pricks usually contain no more than 50ul of blood, and yield approximately 500 ng to 1ug of DNA. This is usually sufficient for PCR analysis. To obtain higher DNA concentrations, elute with 50ul of preheated elution buffer (volumes lower than 50ul greatly reduce yields). Alternatively, the first eluate can be used to perform a second elution.

    相关产品推荐

    全基因合成| 快速DNA/基因合成（不怕高级结构，周期短，价格实惠）

    全基因合成| 快速DNA/基因合成（不怕高级结构，周期短，价格实惠）

    ￥0.80

    SPIN1/SPIN1蛋白Recombinant Human Spindlin-1 (SPIN1)重组蛋白Ovarian cancer-related protein Spindlin1蛋白

    SPIN1/SPIN1蛋白Recombinant Human Spindlin-1 (SPIN1)重组蛋白Ovarian cancer-related protein Spindlin1蛋白

    ￥1344

    KTO: KIT A643 VALIDATION PROTOCOL (L)-Anatel A643a Accessories

    KTO: KIT A643 VALIDATION PROTOCOL (L)-Anatel A643a Accessories

    ￥50094

    糖组学分析| 糖蛋白质组学分析服务| 蛋白质N-糖基化分析| 蛋白质O-糖基化分析

    糖组学分析| 糖蛋白质组学分析服务| 蛋白质N-糖基化分析| 蛋白质O-糖基化分析

    询价

    PROC/PROC蛋白Recombinant Human Vitamin K-dependent protein C (PROC)重组蛋白Anticoagulant protein C;Autoprothrombin IIA;Blood coagulation factor XIV蛋白

    PROC/PROC蛋白Recombinant Human Vitamin K-dependent protein C (PROC)重组蛋白Anticoagulant protein C;Autoprothrombin IIA;Blood coagulation factor XIV蛋白

    ￥1344

    相关问答

    问

    WB显出膜没有条带

    3010 围观
    2019-02-21

    问

    2小时能完成Western Blot实验，可能吗？

    2984 围观
    2019-01-07

    问

    质粒 DNA 比基因组 DNA 更大吗？

    1 回答 2774 围观
    2014-09-12

    相关方法

    质 粒 D N A 的制备

    h E S 细 胞 的 手 工 传 代

    制 备 E B V 转 化 的 B 细胞

    推荐阅读

    E.Z.N.A.® Viral RNA Spin Protocol

    E.Z.N.A.® Mag-Bind® Viral DNA/RNA Spin Protocol

    E.Z.N.A.® Mag-Bind® Viral DNARNA Spin Protocol

    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