Determination of mRNA half-lives in embryonic mouse skin

Embryonic mouse skin samples were halved, one half separated into epidermis and dermis and RNA isolated, and the other half immersed in medium containing Actinomycin D (Act D) to halt transcription. After 30, 60 or 120 minutes at 37 degrees C the Actinomycin D treated samples were separated into epidermis and dermis and RNA isolated from these. Resulting RNA samples were sequenced to quantify transcript abundances.将胚胎小鼠皮肤样本均分为两份：其中一份被分离为表皮（epidermis）与真皮（dermis）组分，并提取核糖核酸（RNA）；另一份则浸没于添加了放线菌素D（Actinomycin D，Act D）的培养基中以阻断转录进程。将经放线菌素D处理的样本在37℃条件下分别孵育30分钟、60分钟或120分钟后，同样将其分离为表皮与真皮组分，并从中提取RNA。对所有获得的RNA样本进行测序，以定量转录本的丰度。