Transcriptomic responses of C.elegans to freeze-thaw stress

To identify genes regulated by freeze-thaw stress (FTS) in C. elegans, we conducted RNA sequencing (RNAseq) of wild-type well-fed C. elegans populations after 25-minute exposure to -20 degree Celsius freezing followed by recovery at 20 degree Celsius for 1 hr. All animals survived under such condition on nematode-growth-media (NGM) culture plates. We used such condition in an attempt to identify genes that specifically and rapidly respond to FTS rather than those that respond to general organismic deterioration, since longer duration of freezing without cryoprotectants or longer recovery causes organismic death.