The solution made in the container was calcium chloride (3% w/v).
E2 was purchased from Sigma Chemical Company (St Louis, MO, USA), with stock solution made in 95% ethanol.
AFDX-116 (Ascent Scientific, Bristol) was used at 1 μM from a stock solution made in DMSO.
After electrophoresis, the DNA in the gel was stained for 30 minutes in GelRed (Biotium, Hayward, CA) solution made in TAE buffer.
After indicated treatment, cells were stained with 0.5% crystal violet solution (made in 25% methanol and stored at room temperature) for 10 min and photographed.
Gradients (10-ml total volume, 30-50%, w:v) were generated from a 50% sucrose solution made in D2O buffered to pH 7.2 with NaOH, and a 30% sucrose solution made in 10 mM Tris-HCl, pH 7.2, 150 mM NaCl, 5.7 mM Na2EDTA.
Briefly, a 0.4 M Mn2+ solution was mixed with 8 M NaOH in 0.5 L of water (both solutions made in deionized/deoxygenated water), which was cooled to 0 °C and oxidized by bubbling oxygen through the solution for 5 h.
The students prepare their own wash buffers from stock solutions (made in lab I).
All drugs used were added to solution E from stock solutions made in dimethyl sulfoxide (DMSO) with an exception of N- 2-naphthalenyl - 3,5-dibromo-2,4-dihydroxyphenyl)methyleN- 2-naphthalenyl - 3,5-dibromo-2,4-dihydroxyphenylN- 2-naphthalenyl - 3,5-dibromo-2,4-dihydroxyphenyl
This is also true of the solutions made in the first day of lab, since many, such as the 5 M NaCl, were used as stocks for critical solutions such as hybridization, wash, and detection buffers.
To determine what the chemical composition was of the resulting WAF solutions made in embryo medium, Solid phase microextraction (SPME,100 mm polydimethylsiloxane) and gas chromatography mass spectrometry (GCMS, Agilent 7890A GC/5975C MSD) were performed.
