For 2mg of Total RNA:

- Wash 200mg of oligo dT cellulose (collaborative research, cat. 20002)
  with 10mL of NETS buffer (100mM NaCl, 10mM EDTA, 10mM Tris HCl pH 8.0, 0.2% SDS)
- Spin down, aspirate, repeat twice
- Resuspend in 1mL 2xNETS
- Add Total RNA in 1 ml
- Mix on a gentle shaker at RT for 1 hour
- Pour slurry onto a fritted funnel over a vacuum flask.
  You can also use disposable BioRad columns (10 ml plastic with frit)
  over a small vac flask using a rubber stopper 
  with a hole in it as a manifold
  with a 125 ml vac flask the spacing is just right so you can collect
  the eluates in 14 ml plastic falcon tubes placed inside the flask
- Wash resin with 1xNETS five times, 3mL each.
- Elute from funnel into a new vacuum flask (or use a new falcon tube)
  with 1mL, 65C prewarmed 1xETS
  buffer (Same as NETS, except no NaCl)
- Repeat 7 times
- Add 1/10th volum 3M NaAcetate, with linear acrylamide as carrier  (20ug).
- Add equal volume of isopropanol, chill to -20C, then precipitate.
- Wash pellet with 70% EtOH.
- Resuspend in water.
- Quantitate with a spectrophotometer

You can also bind in batch and use spin columns:

Oligo-dT cellulose (Ambion #10020 costs $130.00/gram)
empty spin columns from Bio-Rad (#732-6008  $91.00 for 100)

- Use 75 mg of oligo dT cellulose/mg of RNA
- Bind in High Salt
- wash with NETS buffer (100mM NaCl, 10mM EDTA, 10mM Tris HCl pH 8.0, 0.2% SDS)
- elute with 70 C 5 mM Tris pH 7.5