title: GSM8125818: Ctrl ALBSF 04; Mus musculus; RNA-Seq
library_construction_protocol: To collect tissue from the PMBSF and ALBSF territories, ctrl and embWPC P0 pups were euthanized via decapitation, and their brains were dissected out under RNase-free conditions to prevent RNA degradation. The brains (five brains were pooled for each sample) were collected in ice-cold KREBS solution and sliced into 300 μm sections using a vibratome. The PMBSF and ALBSF territories were rapidly microdissected under a stereo microscope. The bulk tissue was immediately transferred to lysis buffer of the RNeasy® Micro Kit (Qiagen, 74004) for total RNA extraction, following the manufacturer's instructions. RNA quality was measured for all samples using an Agilent Bioanalyzer 2100 system, and only samples with RNA Integrity Number (RIN) > 8 were used for library construction. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Then the quantified libraries were  pooled and sequenced for 150pb paired-read and performed on an Illumina Novaseq6000 platform using 2x150 sequencing to read depth of > 40 million reads per sample at Novogene Co. Ltd.  genomics core facility (Cambridge, UK).