title: GSM8703960: Control_10hr; Danio rerio; RNA-Seq
library_construction_protocol: Collected tissue was transferred to 300 uL of digestion buffer (Final concentration .12 mg/mL Liberase-TL (Millipore Sigma, 5401020001) and 22.5 U/mL DNAse I (Worthington, LS006331) in .25\% Trypsin/EDTA (Gibco, 25200-056)) in an eppendorf tube. The tube was placed on a heat block at 30 C for 6 minutes, and a P1000 pipette was used to triturate. The samples were centrifuged at 400 g for 5 minutes and the digestion buffer was removed. Cells were resuspended in 1 mL wash buffer (.1\% BSA in PBS) per tube; filtered through a 40 um cell strainer; centrifuged for 5 minutes at 400 g; and resuspended in 30-50 ul wash buffer. Cell density was estimated by counting on a hemocytometer prior to loading into a 10X Genomics lane. The 10X Genomics Chromium 3' platform and reagents (v3.1 kit) was used for cell encapsulation, reverse transcription, and library preparation, according to manufacturer instructions (manual revision D), with 12 cycles of preamplification. For cell encapsulation, one lane of the 10X chip was used for each treatment group (LPS and control), at a target of 16,000 cells/lane.