Unlike sea urchins, sea stars have little stored histone RNA in their eggs. In an effort to quantify this difference, we have measured H3 RNA concentration in eggs and embryos of the sea star Pisaster ochraceus. The amount of H3 transcript in P. ochraceus 12-h embryos has been measured by RNA excess hybridization kinetics, using a single-strand 32P-labelled coding sequence probe. There are 1 × 105 H3 transcripts in each 12-h embryo. Putative egg H3 transcript concentration was estimated by reciprocal plots. The number of egg H3 homologous sequences (150/egg) is at least an order of magnitude less than rare complex-class, single-copy nuclear DN A transcripts. Slot blots and Northern blots indicate that sea star embryos do not reach the level of H3 transcript abundance seen in sea urchins until at least 16 h of development.
Mixture models reveal multiple positional bias types in RNA-Seq data and lead to accurate transcript concentration estimates