100
ANITA RAUCH
- Fixative:
- Acetic acid/methanol 1:4 (e.g. 50 ml acetic acid plus 150 ml methanol, prepare freshly)
- RNase working solution (l00 ng/I.d): 1 ~l stock solution plus 99 ~l
2xSSC
- Stock solution: dissolve 10 mg RNase A (Roche) in 1 ml 2xSSC,
cook for 10 min and store in portions of 100 ~l in -20°C
- Pepsin stock solution 10%: Dissolve 19 pepsin (Fluka) in 10 ml aqua
bidest and store in portions of 50 ~l at -20°C
2x SSC: 50 ml 20 x SSC plus 450 ml aqua dest
- 20xSSC: 87.65 g NaCl plus 44.1 g Na-citrate plus 500 ml aqua bidest,
adjust pH to 7.0 with HCI 32%, store at room temperature
- PBS: 50 milO x PBS plus 450 ml aqua dest
- lOx PBS: 80 g NaCI plus 2 g KCI plus 11.5 g Na2HP04 plus 2 g KH 2 P0 4
add 1 I aqua bidest, adjust pH to 7.0 with NaOH, store at room temperature
- 1 M MgCh: dissolve 20.3 g MgCh in 100 ml aqua bidest, store at room
temperature
- PBS-MgCh: 10 mIl M MgCl 2 plus 190 ml PBS
- 1% Formaldehyde: 2.7 ml formaldehyde (37%, Merck) plus 97.3 ml
PBS-MgCh
Denaturation - 70% formamide/2xSSCx: per slide: 70 ~l deionized formamide plus
10 ~l phosphate buffer plus 10 ~120x xSSC plus 10 ~l aqua dest (may
be stored at -20°e)
- Phosphate buffer: dissolve 0.7 g sodium dihydrogen phosphate monohydrate in 10 ml aqua bidest, dissolve 0.89 g sodium dihydrogen phosphate dihydrate in 10 ml aqua bidest and mix both, store at -20°C in
portions of 1 mt.
- Deionized formamide: ad 2.5 g ion exchanger (mixed bed resin) to
50 ml formamide, stir for 2 h, let the ion exchanger settle down and
aliquot formamide in 1 ml portions, store at -20°e.