title: GSM5929388: dEmbryo_45min - MP1 antibody m6A-IP-seq; Drosophila melanogaster; OTHER
library_construction_protocol: Timed egg lays were used to stage embryos by adding a fresh molasses agar plate with yeast paste to the collection cage for 30 minutes; this was then replaced with a fresh plate for the next collection cycle. The plate was submerged in 1X PBS and the eggs were dislodged with a paint brush then inverted over a funnel into an egg basket and rinsed with additional PBS. The egg basket was then inverted over a 50 mL conical tube and the eggs were collected with PBS and pelleted for 3 min at 300 x g. The PBS was removed and the eggs were transferred with a wide bore pipette tip in approximately 1 mL of PBS to a 1.5 mL microfuge tube and pelleted for 2 min at 500 x g. The PBS was removed and the egg pellet was flash frozen in liquid nitrogen and stored at -80°C. On average, the embryo pellets were frozen 15 minutes after removal of the agar plate from the collection cage yielding embryos between 15 and 45 minutes of development (referred hereafter as 45min embryos). The pellets were transferred to 1.2 mL of Buffer A supplemented with 1% sodium dodecyl sulfate, 0.5% polyvinylpyrrolidone and fresh 1% 2-Mercaptoethanol, and DNA was extracted as described for cauline leaves.