title: GSM2966348: Long-range VIP-GABAergic cell from the VIP-eGFP mouse hippocampus s3; Mus musculus; RNA-Seq
library_construction_protocol: acute hippocampal slices in ACSF, patch-clamp recordings followed by cell mechanical aspiration, the cell content was extracted in the lysis buffer containing the RNAse inhibitor The cDNA libraries were constructed using SMART-Seq v4 Ultra low input RNA Kit. cDNA from SMART sequences was amplified by PCR Primer II A. After 17 cycles of long-distance PCR, amplified cDNA was purified using the Agencourt AMPure XP Kit.The quality of cDNA library construction was validated using the Agilent Tapestation 2200 system. The threshold cDNA concentration of 200 pg/ml was achieved in all samples. cDNA libraries from 23 cells were prepared for Illumina Next Generation sequencing using Nextera XT DNA Library Preparation Kits. Libraries with unique index were then pooled together in equimolar ratio and sequenced for paired-end sequencing using both lanes of a rapid run flow-cell on the Illumina HiSeq 2500 system. The average insert size for the paired-end libraries was 225 base pairs.