GSM5906711: Mouse 6; Mus musculus; OTHER
The STARR-seq protocol was performed according to the recommendations of Muerdter et al. 2018. After sacrificing the mice, cortical tissue surrounding the injection sites was quickly removed and frozen using liquid nitrogen. Total mRNA was isolated from individual brain tissue samples using the AllPrep DNA/RNA Mini Kit (Qiagen; Cat. No. 80204) according to the manufacturer's protocol (version 2005). Oligo-dT selection of mRNA was performed with NEBNext® Poly(A) mRNA Magnetic Isolation Module (NEB; Cat. No. E7490S/L) with a starting amount of 5µg (except for sample 1 with a starting amount of 2.7µg). Reverse Transcription was performed with SuperScriptIII (Invitrogen; Cat. No. 18080093) using a Gene specific primer (CTCATCAATGTATCTTATCATGTCTG). RT was followed by RNAse Treatment and junction PCR with 15 cycles. Libraries were amplified in 8 Cycles. RNA libraries were prepared for sequencing using standard Illumina protocols.