
The second dissociation step allows the breakdown of any remaining bead-organism complexes and maximizes the number of organisms released from the beads.
- Add another 50 µL of 0.1N HCl to the microcentrifuge tube and repeat the steps of the first dissociation.
The suspension resulting from the second dissociation may be applied to the same slide well as the suspension from the first dissociation or a different slide well.
If using the same slide well:
- Add another 5 µL of 1.0N NaOH to the well either before or after applying the second dissociation to the well.
If using different slide wells:
- Add 5 µL of 1.0N NaOH to a second well.
- Add the second dissociation volume to this well.
Follow Method 1623 and manufacturer instructions from staining kit for specific drying techniques.

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