Transcriptome sequencing on whole embryo of wild-type zebrafish and three bmpr2a/b mutant genotypes (bmpr2a-/-:bmpr2b+/+, bmpr2a+/+:bmpr2b-/- and bmpr2a-/-:bmpr2b-/-) at 48 hpf.
To investigate the molecular mechanisms by which bmpr2a and bmpr2b regulate early zebrafish development, this project used bmpr2a/b heterozygous (bmpr2a+/-:bmpr2b+/-) zebrafish for breeding to generate four experimental genotypes: wild-type (bmpr2a+/+:bmpr2b+/+) and three bmpr2a/b mutant genotypes (bmpr2a-/-:bmpr2b+/+, bmpr2a+/+:bmpr2b-/-, and bmpr2a-/-:bmpr2b-/-). Transcriptome sequencing was then performed on these genotypes following the workflow below:At 48 hours post-fertilization (hpf), tail biopsies were collected from juvenile zebrafish for genotyping, while the remaining embryonic tissues were immediately snap-frozen and stored at negative 80 degrees celsius to preserve RNA integrity. After genotyping confirmation, embryonic tissues from four individuals of the same genotype were pooled into a single sample tube. Total RNA was extracted from each pooled sample, followed by cDNA library construction and RNA sequencing (RNA-seq). For each genotype, three biological replicates were set up, with each replicate containing tissues from six juvenile zebrafish.