Single nuclei RNA sequencing of patient sample: Sample NR1CST
We extracted single nuclei from two specimens (melanoma and NSCLC) using the salt-tris (ST) based extraction method with mechanical dissociation using spring scissors. frozen tissue was placed inside a 6 well plate and 1 ml of either TST buffer (see above) or ST buffer with 0.5 % CHAPS (Millipore, 220201) (CST buffer) were added. Tissues were then chopped for 10 minutes using Noyes spring scissors (Fine Science Tools, 15514-12). 0.9-1.5x103 nuclei were then loaded in ST buffer with or without RNAse inhibitor using a Chromium controller and chromium reagents (10X Genomics) for 3' (#1000075) or 5' capture (#1000006 and #1000263) as indicated.