title: GSM3536476: 3'mRNAseq_zPCF11deltaPAS1_hom_e6; Danio rerio; RNA-Seq
library_construction_protocol: Dechorionated embryos of heterozygous Pcf11-mutant incrosses were cut in half with a razor blade at 19 hpf (zPCF11null) or 32 hpf (zPCF11deltaPAS1) and, and each head and tail was  collected individually in PCR tubes. The anterior halves (heads) were lysed in 10 µl of TCL buffer with 1% beta-mercaptoethanol and flash-frozen on dry ice for subsequent use for RNA isolation and sequencing. The posterior halves (tails) were used for genotyping of each individual sample. RNA of selected samples was isolated and purified using Agencourt RNAClean XP magnetic beads (A63987, Beckman Coulter). Libraries were prepared according to the protocols of QuantSeq 3'mRNA-Seq library prep kit FW for Illumina (LEXOGEN Cat# SKU 015.96) 3-4 wild-type, 4-5 heterozygous, and six homozygous samples of both zPCF11 null and zPCF11 delta-PAS1 mutants were used for library preparation. RNA of selected samples was isolated and purified using Agencourt RNAClean XP magnetic beads (A63987, Beckman Coulter). Strand-specific libraries were generated using the QuantSeq 3' mRNA Library Prep Kit FW (Lexogen) and used for 100-bp single-end sequencing on the Illumina HiSeq 2500.
attributes.tag: GEO Accession
attributes.value: GSM3536476