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        The E.Z.N.A.® Mag-Bind Dye terminator Removal Procedure

        互联网

        950

        实验试剂

         

        1. Absolute (96%-100%) ethanol

        2. Molecular biology grade water

        实验设备

         

        1. Multiple-channel pipettor

        2. 96-well microplate

        3. Magnetic Separation Devices

        实验步骤

         

        1. Determine the volume of the crude sequencing product and transfer the sample into 96-well microplate. Add 10 ul molecular grade water to each sample.

        2. Add 2 volume of Mag-Bind® Particle/ethanol mixture and mix thoroughly by pipetting.

        3. Incubate 10 minutes at room temperature. Place the microplate on a magnetic separation device designed for 96-well plate. Wait for 5 minute or until the solution is clear of beads.

        4. Aspirate entire solution with multiple channel pipettor. Do not disturb the Mag-Bind™ Particle pellet.

        5. Dispense 100ul MPG Wash Buffer into each well, resuspend Mag-Bind Particle pellet by pipetting. Incubate for 1 minute at room temperature.

        6. Place the microplate on a magnetic separation device, and wait 1 minute or until the solution is clear of beads.

        7. Aspirate entire solution with a multiple channel pipettor. Do not disturb the Mag-Bind™ Particle pellet.

        8. Dry the Mag-Bind™Particle pellet by air for 10 minutes.

        9. Remove the plate from separation device. Add 15ul - 30ul water to each sample and resuspend Mag-Bind® Particle pellet by pipetting.

        10. Incubate 10 minutes at room temperature. Place the microplate on a magnetic separation device designed for 96-well plate. Wait for 1 minute or until the solution is clear of beads.

        11. Transfer the cleared supernatant to a new microplate (not provided).

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