Hybridization Bags Troubleshooting

Product No. 11666649001

Troubleshooting

Background Issues:

High background is caused by non-uniform distribution of chemiluminescent substrate, due to incubation of membrane wrapped in plastic wrap. Wrinkles in the hybridization bag can cause uneven contact between membrane and X-ray film. Background can also be increased by the drying of membrane during the chemiluminescent visualization procedure.

Grainy appearance of the background indicates that drying occurred during the chemiluminescent procedure rather than during hybridization. Drying during hybridization leads to a cloudy background .

Recommendation:

Do not wrap membrane in plastic wrap during incubation with chemiluminescent substrate. Use Roche Hybridization Bags. Spread the chemiluminescent substrate uniformly over the surface of the membrane. Before exposing bag to X-ray film, flatten any wrinkles between blot and membrane by rolling a pipette over the surface of the bag.

Carefully seal the damp membrane in a folder or bag during the incubation/exposure to X-ray film. Check the seals to make sure liquid cannot leak.

Materials

     
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