Northern Blot Oligonucleotype Hybridization

1. Prewash blot in 0.1X SSC/0.1% SDS for 1 hour at 65ºC.
2. Remove prewash.
3. Prehyb blot for >1 hour in pre-hybridization solution at hybridization temperatuer (see below).
4. Remove pre-hybridization buffer.
5. Add probe to blot. Hybridize for >6 hours, at least 15ºC below estimated T_m.
        T_m(ºC) = 4(G + C) + 2(A + T)
6. Pour probe off of blot. Store probe at -20C.
7. Wash blot in 6X SSC, 0.1% SDS
   Three five minute room temperature washes.
   One 20 minute wash at 10ºC below estimated T_m.
8. Dry blot by pressing between paper towels.
9. Wrap blot in Saran Wrap.
10. Expose on film or PhosphorImager cassette.