Name: Zebrafish embro 1 dpf mRNA 3' end
Instrument: Illumina Genome Analyzer II
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: other
Layout: PAIRED
Construction protocol: 20 ug of total RNA was fragmented using RNA Fragmentation Reagent (Ambion) for 5 minutes at 70 C and ethanol precipitated with glycogen and LiCl. RNA was annealed to the oligo stBPM1polyT22 (biotin-GGCCAGTCCTGGAGTTTTTTTTTTTTTTTTTTTTTTVN) and bound to streptavidin magnetic beads. After washing by pull down on a magnet, the bound RNA was reverse transcribed with SuperScript II (Invitrogen) and a second strand synthesised with DNA polymerase I (Promega) and RNase H (NEB). After further washing, the double strand cDNA was released from the beads with BpmI (NEB). The cDNA was recovered with the QIAgen PCR Purification Kit and made into a standard Illumina library following the manufacturer's protocol with a fragment size of 250 to 300 bp.