I am getting high DAB background staining. How can I decrease this background in the future?

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Answer

High DAB background can be decreased by taking the following steps:
1) Make sure that the stringent wash is performed properly.
2) After hybridization, rinse the slides briefly in room temperature SSC buffer, then immerse slides for 5 minutes in SSC at 75°C.
3) Increase the temperature by 1°C per slide for more than 2 slides, but do not exceed 80°C.
4) For any other wash steps , 1X PBS with Tween® 20 (0.025%) is recommended. Washing with any other reagents (1X PBS without Tween® 20 or distilled water) can lead to unwanted background.

Answer Id: E5005

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