WASH PROTOCOLS:
SSC HYBS:
Dismantle wash: 1x SSC, 0.1% SDS
Wash 1, at hybridization temperature: 1x SSC, 0.1% SDS 10 min.
using the Variomag submersible stir plate and a beaker in a waterbath.
(See Donna or Jessica for demonstration)
Wash 2 RT: 1x SSC 10 min.
Wash 3 RT: 0.1x SSC 5 min.
Spin dry and scan.
If you find you are still getting streaky arrays, add wash 4, 0.01xSSC 30s.
AGILENT HYBS:
Done with stir bars in glass dishes, maximum 6 slides per batch
Disassembly wash (same as wash A)
Wash A (1 L) 1min
Add in this order:
700 ml water
300 ml 20X SSPE
0.25 ml 20% N-lauroylsarcosine
Filter. Shake to mix.
Wash B (1 L) 1min
Add in this order:
997 ml water
3 ml 20X SSPE
0.25 ml 20% N-lauroylsarcosine
Filter. Shake to mix.
Wash C: 30s Remove slowly to avoid streaks.
HPLC grade Acetonitrile