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文献和实验Single and Double FISH protocols for Drosophila
3 transcription optimized buffer (Promega, Madison WI, USA; Cat No. P1181) 4. T7 or T3 RNA Polymerase (Promega, Madison WI, USA: Cat Nos. P2075, P2083; 1000 U). 5. Fluorescein RNA labeling Mix (Boehringer Mannheim; Cat No. 1685 619). 6. Digoxigenin RNA
In Situ HYBRIDIZATION TO TISSUE SECTIONS
ml glass tube -Spot 10ml on a GF/C filter disk -To the remaining 90ml, add 5ml cold 10% TCA; incubate 10' on ice -Filter onto GF/C, wash with 10% TCA, wash with 95% EtOH and dry -20-50% incorporation is good and corresponds to a S.A. ‰ 1-2 x 108
by tapping the tube (try to avoid getting beads stuck to the side of the tube, above the level of the reaction mixture). 14.) Incubate at 25oC (room temp.) for 25min, mixing occasionally. 15.) Add sample buffer to 1x (I usually add 20 μl of 3x
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