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文献和实验RNA Whole Mount In Situ Hybridization
, but it could probably be used successfully. Wash 10 minutes in a 1:1 mixture of hybridization solution/PBT Wash 10 minutes in hybridization solution. Incubate at 70°C in hybridization solution for at least 1 hour. Replace hybridization solution
Whole mount in situ hybridization with digoxygenin probes
: PBT = 1:1 5' with 100 µl Hyb sol Prehybridize with 100 µl Hyb sol at 45 - 50 C > 1 hr. Denature probe by boiling 5'; chill on ice 3' Hybridize with probe in 50-100 µl Hyb. sol at 45 - 50 C O/N. 7. Washes SAVE THE PROBE
(either undiluted or 1:1 with PBT) or diluted polyclonal antiserum and incubate at 4 C overnight 8. Rinse with PBT 3 times, then wash 4 times for 15 minutes each 9. Add secondary antibody (usually 1:200 in PBT) and incubate 2 hours at room temperature
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