title: GSM8686971: LAG3+ NK cells Day2 replicate 1; Mus musculus; RNA-Seq
library_name: GSM8686971
library_construction_protocol: RNA was harvested using GenElute™ Mammalian Total RNA Miniprep Kit (Sigma Aldrich) The entire process of experiments from library preparation to sequencing was performed by DNA Link (DNA Link Inc., Seoul, Republic of Korea). mRNA was purified and fragmented from total RNA using poly-T oligo-attached magnetic beads using two rounds of purification. Cleaved RNA Fragments primed with random hexamers were reverse transcribed into first strand cDNA using reverse transcriptase, random primers, dUTP in place of dTTP. These cDNA fragments then had the addition of a single 'A' base and subsequent ligation of the adapter. The products were purified and enriched with PCR to create the final strand specific cDNA library. The quality of the libraries was verified by Tapestation (Agilent Technologies, Santa Clara, CA, USA). After quantitative PCR (qPCR) using KAPA SYBR FAST qPCR Master Mix (Kapa Biosystems, Wilmington, MA, USA), we combined libraries that index tagged in equimolar amounts in the pool.