mRNA-Seq: adult SA male rat ventral tegmental area
The concentration and quality of purified total RNA was determined on Qubit 3.0 and by Agilent 2100 Bioanalyzer, respectively. Total RNA libraries were performed using VAHTS Universal V8 RNA-Seq Library Prep Kit for Illumina, respectively. 300 ng total RNA was used. First, RNA was incubated with rRNA probe to form rRNA-probe hybrid, which was depleted by RNase H. Then the probe was removed by DNase I digestion. Then the rRNA depleted RNA was subjected to fragmentation. Strand-specific library was prepared by inclusion of dUTP in cDNA synthesis and digestion with UDG. Size selection was performed to selectively enrich insertion size range from 150 bp to 200 bp. The sequencing was performed using NovaSeq 6000, by multiplexing samples with dual barcoding index and the sequencing depth was 50 million reads on average.
F1-SA-CSA_ventral tegmental area_rep_2