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Karyotyping: from microscope to array - II
Stripping Agilent Arrays For Reuse
1. Use a slide staining glass reservoir plus slide rack and lid. A handle attached to the rack will prevent burnt fingers. Fill the reservoir with 50:50 Agilent Wash Buffer2: Hplc water
2. Set a heater/stirrer to 65°C and allow to equilibrate. Microwave the reservoir with wash buffer2/water to 65°C. Don't put the cold reservoir on the hot plate as it cracks. Add a magnetic stirrer
3. Scan the array to record the maximum intensities in Feature Extraction (select ‘set colour range’ on the tool bar) before stripping. Place the array into the reservoir with magnetic stirrer for 45min. Strip up to 3 arrays at a time. The temp will fluctuate ~5°. After 45 min remove the array slowly (about 10 sec) keeping the array horizontal to permit drying.
4. Rescan array. If the intensities are less than 10% of the intensities before stripping then consider it a pass. Otherwise repeat for another 45 min with fresh buffer and check the intensities again
5. Wash the reservoir carefully after use - about 10 rinses with RO water, 1 rinse in RO water warmed to 37°C (3min high microwave), then a final rinse with hplc water. Absolutely no detergent