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文献和实验96-well RNA In Situ Hybridization Protocol
+ 5% Normal Goat Serum + anti-digoxigenin Alkaline Phosphatase (1:2000 dilution), rock 2 hrs at RT. 20. Rinse twice with PBT . 21. Wash embryos 9 X for 10 min. in PBT. 22. Rinse twice with 200ul Alkaline Phosphatase buffer (AP buffer). 23
96-well RNA In Situ Hybridization Protocol
. at RT. 19. Remove PBT, add 200μlPBT + 5% Normal Goat Serum + anti-digoxigenin Alkaline Phosphatase (1:2000 dilution), rock 2 hrs at RT. 20. Rinse twice with PBT . 21. Wash embryos 9 X for 10 min. in PBT. 22. Rinse twice with 200μlAlkaline Phosphatase
. Add 1ml lysis bufferb and vortex briefly. 11) Repeat Step #10 2-3 more times. 12) After last wash, aspirate supernatant and remove remainder of wash buffer with fine gauge (>22g) needle. Process for kinase reaction (as indicated) or add 40-80μl 1X
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