GSM4232728: IL08_Pool3; Mus musculus; RNA-Seq
P7 E13.5 born Cortical neurons where captured by preparing 300um thick acute coronal brain slices maintained in aCSFcontaining 119 mM NaCl, 2.5 mM KCl, 1.3 mM MgCl2, 2.5 mM CaCl2, 1.0 mM NaH2PO4, 26.2 mM NaHCO3 and 11 mM glucose, bubbled with 95% O2 and 5% CO2. Slices were maintained 30 minutes in bath at 32°C and then at room temperature. The whole-cell voltage-clamp recording technique was used (30–32 °C, 2–3 ml min−1, submerged slices) to measure the holding currents and Ih current of GFP+ FlashTag neurons. The internal solution contained 140 mM KCH3O3S, 4 mM NaCl, 2 mM MgCl2, 0.2 mM EGTA, 10 mM HEPES, 3 mM Na2ATP, 5 mM sodium creatine phosphate, 0.33 mM GTP and 1µl of 1U/mL of RNase inhibitor (Takara). After recording the cells were acquired by manual suction until visual confirmation of the capture of the cytosolic volume. cDNA libraries were prepared for sequencing using standard Illumina protocols using SMART-Seq v4 3' DE kit (Clontech, # 635040) and sequenced using HiSeq 4000 in 100 pairbase length fragments for a minimum of 1 million reads per sample.
GEO Accession
GSM4232728