Homo sapiens
Protocols: For 2D, 75,000 sgRNA-infected C3 MCF10A cells 818 92 were plated into four 12-well plates (Sarstedt 83.3921). The cells were grown for 24 hours and scraped into 100ul of RNA Lysis buffer (Zymo Research R1051), then stored in -80C prior to RNA extraction. For the 3D sphere formation assay in RMRP FMRE experiments, 15,000 MCF10A cells were plated on Matrigel-coated 24-well plates and grown in MCF10A Assay Media with regular media changes every other day for three weeks. After sphere formation, media was removed, cells were rinsed with cold PBS, spun down, and resuspended in Zymo Lysis buffer for RNA extraction. RNA samples were extracted from cells using the Zymo Quick-RNA Microprep Kit (Zymo Research R1051). After extraction, sample integrity was checked with RNA Qubit (Invitrogen Q32852) and the Bioanalyzer.
replicate
2
disease
non-malignant
organism
Homo sapiens
organism
Homo sapiens
collection date
not collected
scientific_name
Homo sapiens
common name
human
organism part
mammary gland
developmental stage
adult
genotype
N/A
isolate
not applicable
sgrna
Scrambled
2d or 3d
3D
cell_line
MCF 10A
cell_type
epithelial cell
geographic location (country and/or sea)
not collected