Hybridization Ovens
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81-0169-01 Rev Q
Non-Radioactive Probes
1. Wash blots in 2X SSC, 0.1% SDS for 3 minutes at room temperature (repeat one)
2. Wash filter in 0.15X SSC, 0.1%SDS for 15 minutes at 50 °C (repeat once)
3. Store blots in 2X SSC at room temperature
Radioactive Probes
Additionally you will need:
•
X-Ray film holder
•
X-Ray film
•
Intensifying screen
Procedure
1. Remove blot from hybridization tube and transfer to Tupperware container
2. Rinse briefly in 50 °C 0.1X SSC, 0.1% SDS.
3. Remove this solution to radioactive waste and wash
4. Wash blot three more times in the same solution.
5. After the final wash, dry blot on filter paper for 10 minutes. This is a good time to quickly pass your
hand held radioisotope reader (beta or gamma counter) over your blot to get a general idea as to
the exposure time you will need for the x-ray film. Hot blots are 20 minutes to 2 hours. Not so hot
blots can be left overnight (8 hours).
6. Tape the blot to a cardboard backing.
7. Cover with plastic wrap to prevent the blots from sticking to the x-ray film.
8. Place the cardboard containing the blots into the X-ray film folder.
9. In the darkroom, place a piece of X-ray film over the blot(s).
10. Place an intensifying screen on top of the film.
11. Close the film folder and clamp it.
12. Store at –70 °C. The low temperature reduces light scattering and increases the length of
exposure time. Expose the blot for 20 minutes to 24 hours.
