Illumina HiSeq 2500 sequencing; CAGE-seq of zebrafish cells extracted at the 14 somite stage and segregated by cell cycle dynamics (e.g. fast - green vs slow - red)
CAGE-seq of zebrafish cells extracted at the 14 somite stage and segregated by cell cycle dynamics (e.g. fast - green vs slow - red)
Zebrafish were dechorionated and selected at the 14-somite stage (14 hours post fertilisation). The embryos were dissociated into a single cell suspension using enzyme-free cell dissociation buffer, PBS based (Gibco). Dissociated cells were pelleted and resuspended in Hanks balanced salt solution without calcium chloride or magnesium sulphate (Sigma), for sorting. Cells were fluorescence-associated cell sorted (FACS) into populations displaying red and green fluorescence. RNA was extracted from isolated cells using the miRNeasy kit (Qiagen). RNA quality was analysed by capillary electrophoresis (Bioanalyzer 2100, Agilent). All samples had an RNA integrity number (RIN) >9. NanoCAGE libraries were generated following a protocol described in:  Poulain, S., Kato, S., Arnaud, O., Morlighem, J.E., Suzuki, M., Plessy, C. and Harbers, M. (2017) NanoCAGE: A Method for the Analysis of Coding and Noncoding 5'-Capped Transcriptomes. Methods Mol Biol, 1543, 57-109.
Experimental Factor: fraction
S/G2/M (rapid cycling)