Illumina HiSeq 1500 sequencing; Impact of RNA isolation methods for RNA-seq on gene expression. Here RNA was isolated from mouse striatal tissue using either Columns or magnetic beads and the isolated RNA was processed using prime-seq.
Impact of RNA isolation methods for RNA-seq on gene expression. Here RNA was isolated from mouse striatal tissue using either Columns or magnetic beads and the isolated RNA was processed using prime-seq.
The brain was transferred to a dish with ice-cold PBS and placed in a 1 mm slicing matrix. Using steel blades, 5 coronal incisions were made. Biopsy punches were then taken from the striatum and the tissue was transferred to a 1.5 mL tube with 50 µL of lysis buffer, RLT Plus and 1% ß-mercaptoethanol. The tubes were snap frozen and stored at -80 ºC until needed.   RNA was isolated using the bead extraction described in the prime-seq protocol The prime-seq protocol was performed as described here: https://www.protocols.io/view/prime-seq-s9veh66