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In short, larvae were fixed in 4% paraformaldehyde/0.05% glutaraldehyde in MOPS buffer and stepped into 70% ethanol.
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Fixed samples were then rinsed several times with PBS buffer plus 0.1% Tween 20 and stepped into 75% ethanol and stored at -20°C.
Once thoroughly decalcified, the material was washed in 1× PBS-Tween and stepped into hybridization buffer (0.75 M sodium chloride, 75 mM sodium citrate, 5 mM EDTA, 50% formamide, 50 μg/ml heparin, 0.1% Tween-20, 0.1 mg/ml yeast total RNA).
Springsteen glanced at the step and stepped into the spotlight.
And step into life.
Novikov and Kolesnikov stepped into the corridor.
Joe Torre stepped down and recently stepped into the Hall of Fame.
Afterwards, larvae were stepped into 100%% hybridization buffer and pre-incubated at 60 °C in a water bath overnight.
Developmental stages were fixed in 4 % paraformaldehyde in MOPS buffer (0.5 M Mops, 10 mM MgSO4, 5 mM EGTA, 2.5 M NaCl) for 45 min at room temperature, stepped into 100%% methanol and stored in 100%% methanol at −20 °C.
Then Dodge stepped into the shadow of his fire, so that he was surrounded by a buffer of burned land.
And I won't step into it.
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