title: RNA-seq of human intratumoral CD14 cells: Sample 562-iCD14-2
design:  LCM harvest protocol : cryosection were fixed in cold acetone and briefly air dried, immunofluorescence staining was performed on a HistoGene cold bloc (ThermoFisher) using modified immunofluorescence staining protocol with higher concentration of conjugated antibodies, all  reagents included SUPERase inhibitor (ThermoFisher). After staining, samples were dehydrated  in 75%, 95% and 100% Ethanol successively, the incubated 5 min in Xylene and air dried. Laser  capture microdissection was performed on an Arcturus XT LCM with CapSure Macro LCM caps ThermoFisher). After harvest, RNA was isolated with PicoPure RNA isolation kit following manufacturer protocol. SMART-Seq V1/V4 Ultra Low Input RNA Kit was used for cDNA synthesis and amplification. cDNAs were processed to the library preparation with NEBNext DNA Library Prep Master Mix Set for Illumina and 75bp single reads were used on Illumina NextSeq500 or Novaseq sequencer (Illumina).