Library |
Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
Planarian worms were placed at center of cryomold emerged in ice and water mixture to keep worm stational before applying chilled cryostat (O.C.T. Compound) embedding media (Sakura, 4583), followed by immediate snap-freeze in isopentane and liquid nitrogen bath. OCT embedded sample blocks were cryosectioned in a pre-cooled cryostat at 10 µm thickness and placed onto the active capture area of Visium TO and GE slides (10x Genomics, Visium). Permeabilization time of 22 minutes were determined using TO slide according to manufacturer’s instructions. GE slides were fixed and H&E stained before imaged by Perkinelmer Polaris at 40x magnification and Leica Aperio CS2 at 20x magnification. The tissue was then permeabilized, and reverse transcription and second-strand synthesis were performed, followed by library construction according to the manufacturer’s instructions. Libraries were sequenced on an Illumina Nova platform using 150 base-pair paired-end dual-indexed set up (High output, v 2.5, Illumina). |
RNA-Seq |
TRANSCRIPTOMIC |
unspecified |
PAIRED
|
|