When a large number of C. elegans need to be synchronized, prepare 2-3 plates where more than 80% of the insects in the plates are in the reproductive period, and proceed as follows: (1) Take 5 ml of M9 buffer to wash the plates twice, suck the M9 buffer containing insects into a 10 ml centrifuge tube, centrifuge at 1200 rpm for 3 min, and discard the supernatant; (2) Add 5 ml of freshly prepared synchronized bleaching solution, shake vigorously at room temperature for 3 min to corrode the adult insects, and tiny eggs can be seen under the microscope. Centrifuge at 1200 rpm for 2 min, and discard the supernatant; (3) Add 5 ml M9 buffer to suspend the precipitation, centrifuge at 1200 rpm for 2 min after mixing, discard the supernatant, wash 4 times; (4) Centrifuge for the last time, leave about 2 ml of the solution when discarding the supernatant, and shake well; (5) Pour the solution into a petri dish that has not been inoculated with OP50 and cultivate at 20 ℃ for 18-24 h. (Ozpinar 2020) .
Transfer the larvae to a growth plate (10-20 µL per plate) to obtain synchronized C. elegans.