title: GSM7321056: Mcpip1 DN_replicate1; Danio rerio; RNA-Seq
library_construction_protocol: Zebrafish embryos were collected in Fenozol (A&A Biotechnology, Gdansk, Polska) and stored at -80°C  prior to analysis. For RNA isolation, the samples were homogenized in Fenozol using a tissue homogenizer (OMNI International, Kennesaw, GA, USA). The poly (A) mRNA fraction from total RNA was isolated with Dynabeads® mRNA DIRECT™ Micro Kit (Thermo). The sequencing library for each RNA sample was prepared according to the protocol provided by the manufacturer using the Ion Total RNA-Seq Kit v2 (Thermo). The libraries were generated from 1-15 ng of mRNA by fragmenting the mRNA with RNaseIII, purifying the fragmented RNA, and hybridizing and ligating it with Ion adaptors. Subsequently, the RNA products were reverse transcribed and amplified to double-stranded cDNA, and then purified using a magnetic bead-based method. The molar concentration and size of each cDNA library was determined using the DNA HS Kit on Bioanalyzer 2100 (Agilent). Each library was diluted to ~53 pM concentration before template preparation. Up to three barcoded libraries were mixed in equal volume and used for automatic template preparation on the Ion Chef instrument (Thermo) using reagents from the Ion PI Hi-Q 200 Kit (Thermo) and Ion PI v3 Proton Chip.