1. Add about 20 ml 2X SSPE/0.1% SDS to the tube. Mix and pour out. Be careful of loose drops.
2. Fill tube ≈ 1/3 -1/2 full with 2X SSPE/0.1% SDS and incubate at 56℃ for 30 min.
3. Repeat with 1X SSPE/0.1% SDS and 0.1X SSPE/0.1% SDS until background counts are below 1000 cpm.
        Keep washing with 0.1X SSPE/0.1% SDS if the counts are high. Washes can go longer than 20 min.
        The stringency of the washes depends on the nature of the probe.
4. Wrap the blot with Saran wrap making sure there are no wrinkles.
5. Expose to autoradiography film overnight or to a Molecular Imager screen for 1 hr.
        Adjust exposure time accordingly. The time on the screen is roughly one-tenth to what it would be on film.

   SOLUTIONS

   GITC buffer (200 ml):
   94.53 g Guanidine isothiocyanate (4 M final)
   1.67 ml 3 M NaOAc, pH 6
   Bring volume to 200 ml
   Add 1.67 ml ß-mercaptoethanol
   Store a 4℃ and away from light
   CsCl buffer (100 ml):
   95.97 g CsCl (5.7 M final)
   0.83 ml 3 M NaOAc, pH 6
   Bring volume to 100 ml
   Sterile filter with 0.8 µm
        (This is optional.)
   10X MOPS:
   0.4 M 3-[N-Morpholino] propanesulfonic acid, pH 7.0
   0.1 M Sodium acetate
   10 mM EDTA (pH 8.0)
   Prepare in DEPC water. OK if solution turns yellow.
   Autoclave to sterilize.
   DEPC water:
   Add DEPC to ddH₂O at 1:1000 dilution
   Let it sit overnight at room temp (destroys RNase)
   Autoclave the next day (destroys DEPC)
   RNA sample buffer (7.25 ml):
   0.5 ml 10X MOPS
   1.75 ml 37% Formaldehyde
   5.0 ml Formamide
   Keep away from light!
   Store at -20℃
   RNA loading buffer:
   50% Glycerol
   1 mM EDTA
   0.25% Bromophenol blue
   0.25% Xylene cyanol
   Use DEPC water, keep at 4℃
   2X Slaman hybe:
   2% SDS
   2 M NaCl
   Will not be in solution at RT
   Keep at 4℃
   20X SSPE:
   3.6 M NaCl
   0.2 M NaH₂PO₄-H₂O
   0.02 M EDTA
   pH to 7.7

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