Category Archives: Protocols

RNA in situ hybe buffers

Hybe Solution 50% Formamide (Amresco, ultrapure) 5x SSC 100 ug/mL salmon sperm DNA 50 ug/mL heparin 0.1% Tween-20   Reagent For 250 mL stock Formamide 125 mL 20x SSC 62.5 mL Heparin 10 mg/mL lab aliquot 1.25 mL 10% Tween-20 … Continue reading

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RNA in situ hybridization

 Day 1 Xylene clearing ~2 hrs Rock embryos in 1:1 MeOH:EtOH 5 min (Waste -> organic waste bottle) Rinse 2x with EtOH Rock in EtOH 5 min Remove all but 100 uL EtOH, Add 900 uL Xylenes 1.0 hr (embryos should … Continue reading

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synthesis of RNA in situ probes

Set up in vitro transcription reaction as follows: Reagent Volume 5x transcription buffer (ProMega): 4ul 10x hapten (Dig. Bio DNP) U-NTP mix 2ul 100mM DTT 1ul RNase inhibitor (RNasin) (50 μ/μl) 2ul linearized DNA (~250 ng if 0.2μg/μl) 1ul (vary … Continue reading

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