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        E.Z.N.A. Mag-Bind Blood DNA Protocol (1-100 ul Blood)

        互联网

        1374

        实验试剂

         

        Regents to be provided by user

        1. Absolute ethanol (96%-100%)

        实验设备

         

        Equipments to be provided by user

        1. Nuclease-free 1.5 mL centrifuge tube (for single tube preparation)

        2. 500 ul Nuclease-free microplate

        3. 1.2mL deep well plate (EV1772)

        4. 8-strip caps for 1.2mL deep well plate (EV1774)

        5. Water bath, incubator or heating block preset at 65° C

        6. Magnetic separation device for microcentrifuge tubes or microplates

        实验步骤

         

        1. Add 1-100 ul blood sample to a nuclease-free microcentrifuge tube. Bring the volume up to 100 ul with Elution Buffer provided with this kit.

        2. Add 100 ul Buffer MSL and 10 ul Proteinase K to each sample. Mix by vortexing at maxi speed for 30 seconds.

        3. Incubate sample at 65°C for 15 min. Briefly vortexing the tube during incubation.

        4. Add 150 ul absolute ethanol and 10 ul Mag-Bind Particles Solution C to the sample. Mix by vortexing at maxi speed for 30 seconds.

        5. Place the tube on a magnetic separation device to magnetize the magnetic particles. Incubate at room temperature for 10 minutes.

        6. Completely remove and discard the cleared supernatant. Remove any droplets of liquid from the wall of the tube.

        7. Remove the tube containing the Mag-Bind particles from the magnetic separation device. Add 300 ul Buffer MP/Ethanol mixture to each sample.

        8. Resuspend the Mag-Bind particles pellet by vortexing at maxi speed for 30 seconds.

        Complete resuspension of the Mag-Bind particles pellet by pipetting up and down or vortexing is critical to obtain good results.

        9. Place the tune onto the magnetic separation device to magnetize the Mag-Bind particles. Completely remove and discard the cleared supernatant.

        10. Remove the tube containing the Mag-Bind particles from the magnetic separation device. Add 300 ul SPM Buffer to each sample.

        11. Resuspend the Mag-Bind particles pellet by vortexing at maxi speed for 30 seconds.

        12. Place the tube onto the magnetic separation device to magnetize the magnetic particles. Completely remove and discard the cleared supernatant.

        13. Remove the tube containing the Mag-Bind particles from the magnetic separation device. Add 300 ul SPM Buffer to each sample.

        14. Resuspend the Mag-Bind particles pellet by vortexing at maxi speed for 30 seconds.

        15. Place the tube onto the magnetic separation device to magnetize the magnetic particles. Completely remove and discard the cleared supernatant.

        16. Leave the tube to air dry on the magnetic separation device for 10 minutes. Remove any residue liquid with a pipettor.

        17. Remove the tube from the magnetic separation device. Add 50-100 ul Elution Buffer to each sample. Resuspend the Mag-Bind particles by vortexing at maxi speed for 30 seconds.

        18. Incubate at 65° C for 10 min. Resuspend the Mag-Bind particles by vortexing at maxi speed for 30 seconds.

        19. Place the tube onto a magnetic separation device to magnetize the Mag-Bind particles.

        20. Transfer the cleared supernatant containing purified DNA to a new 1.5 mL tube or plate.

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