LPS Anti-Body Alpha 7 Nicotinic Receptor Treatment
The whole brain was dissected and rapidly frozen in liquid nitrogen stored at -80 degrees. Total RNA was extracted using TRI Reagent (Sigma-Aldrich) according to manufacturer’s directions. Libraries for next generation sequencing (NGS) were prepared from whole brain RNA using TruSeq Small RNA Library Prep Kit as per the manufacturer’s protocol. A total of 4 libraries (pooled from 4 animals of each group) were prepared from RNA of 4 four groups. The cDNA libraries were amplified using bar coded primer sets and 15–18 cycles of PCR. The amplified cDNA libraries were cleaned up and size selected from gel—PCR products of 105–150 bp were isolated, corresponding to inserts derived from the small RNA population. The amplified cDNA libraries generated were there after used for Miseq sequencing.