Nonradioactive in situ hybridization to sectioned tissues

~JECHNICAL

~I~IPS

work well in the alkali blot procedure2; Hybond-N and Nytran gave poor results, and transfer could not be improved by prolonged blotting time or by changing the molarity of the NaOH solution to 0.25 M or 0,6 M, The modgied !dry blot'

[

!

1 RNA

hb

Kr

e

~!es~,thus of

a l l y i n g a !comparison expression pat, 3350 2:1

se~ions We e m ~ d in ~ s ~ e Tek (Miles), freeze in l~uid nitrogen and sectiem at ~lS°C. ~ e sections (8 tim)

slides, The s a

protein

protein

FIGH Comparison of RNA (top) and protein (bottom) expression patterns of the segmentation genes hunchback (hh) (left) and Kr~ppel (Kr) (right) in parallel sections of early Drosophila embryos. The secondary hunchback pattern 1 becomes apparent at a time when the protein expression is still homogeneous. For Kr~ppel it is evident that the protein domain is broader than the RNA domain.

!igestion f°r the in situ hybridization ~ r i m e n t s . However, we d o not dehydrate in n o t d ~ before the ~ining is completedl ~Aand staring for seveol h ~ r s or overnight

~RENCES

1 Tautz, D. and Pfe~e, C ( 1 ~ ) Ch~mosoma 98, 8145 2 IMgoxygenin Application Manual (I989) Boehringer Mannheim, FRG 3 Brown. R.C., ~ m m o n , B.E. and Mullinax, J.B. (1989)BoL Acta 102, 5 ~ 1 Contributed by Ralf ~ m e r and Diethard Tautz, Institut~ r Genetik und mikrobiologie, UnivemitditMf~nchen maria-Wardstr l a, ~ manchen 19, FRG.

TIG AF'RIL1991 rOE. 7 ~O. q l i(i