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Home › Archived Educator Resources › Sanger Sequencing Archive › How Sanger Sequencing Was Done › Step 9: Post-Sequencing-Reaction Cleanup

Step 9: Post-Sequencing-Reaction Cleanup

We’re almost ready to read the bases by capillary electrophoresis. But first the reaction must be cleaned up. To remove the unused product of the sequencing reaction,

oneBeads (magnetic microparticles) are added to the sequencing reaction solution. DNA, but not the excess dye terminators, is induced to attach to the beads.

two Magnets draw down the DNA and beads so the waste solution can be aspirated.

three The beads are washed with ethanol and then dried.

four Water is then added to the beads, and the DNA strands detach from the beads and reenter solution because they have higher affinity for water than for the beads.

five Magnets are then used to secure the beads while the pipettor removes the clean DNA/water solution to a new plate.

cleanup

  • How Sanger Sequencing Was Done
    • Step 1: Shearing of DNA
    • Step 2: Insertion of Fragments into a Plasmid
    • Step 3: Transformation
    • Step 4: Sub-Cloning the Sheared Fragment
    • Step 5: Colony Picking
    • Step 6: Lysing the Cell
    • Step 7: Rolling-Circle Amplification
    • Step 8: Sequencing Chemistry
    • Step 9: Post-Sequencing-Reaction Cleanup
    • Step 10: Capillary Sequencing
    • Step 11: Assembly
    • Step 12: Quality Assessment

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