title: NextSeq 500 sequencing; Tumor metastasis to lymph node requires YAP-dependent metabolic adaptation
design: Tumor metastasis to lymph node requires YAP-dependent metabolic adaptation
library_construction_protocol: 5 x 10^5 B16F10-GFP cells were implanted subcutaneously into the footpad region of hind limb of 8~9-week-old male C57BL/6 mice. B16F10-GFP+ tumor cells from primary site (footpad) and pLNs (isolated at 3 weeks after implantation for micro-metastatic tumor with metastatic foci diameter less than 2 mm and at 4 weeks after implantation for macro-metastatic tumor) were isolated.  Tissues were removed, digested, depleted CD45+ cells using AutoMACS, and GFP+ tumor cells were sorted by FACS Aria II. RNA was harvested using Trizol reagent (Invitrogen). 500 ng of total RNA was used for the construction of sequencing libraries.  RNA libraries were prepared for sequencing using LEXOGEN Quant-Seq Library Prep Kit (Cat#001.24) standard protocols.
attributes.tag: Experimental Factor: disease
attributes.value: LN macro-metastatic tumor