small RNA sequencing of the human hippocampus (adult male)
Small RNAs (15-40 nt) were isolated from a denaturing polyacrylamide gel. Subsequently, a 3 adapter (5-rAppCTGTAGGCACCATCAATddC-3) and a 5 adapter (5- GACUGGAGCACGAGGACACUGACAUGGACUGAAGGAGUAGAAA-3) were ligated to the small RNAs. Following cDNA synthesis with the RT-Primer 5-ATTGATGGTGCCTACAG-3, the cDNA transcripts were PCR amplified using the forward primer 5-ACATGGACTGAAGGAGTAGA-3 and the index-containing reverse primer 5-ggctcATTGATGGTGCCTACAG-3. The generated library was high throughput sequenced in parallel with six other indexed samples on an Illumina HiSeq 2000 system.