Homo sapiens
Protocols: Cells were treated with 2 μg/ml doxycycline and 0.06 μg/ml nocodazole, 24 and 18 h respectively, before the addition of 1 mM IAA to induce degradation of mAID-tagged proteins. After 4 h treatment with IAA, mitotic cells were manually detached by shake-off, and cells were transferred to new poly-L-lysin coated dishes. Once attached, cells were washed 3x with PBS and released from mitosis into G1 for 18 h in a fresh medium containing 2 mM thymidine.Cells were washed with PBS, detached with TrypLE and pelleted by centrifugation. Total RNA was extracted from HCT116 cells (G1, -/+ mAID-protein of interest) using Monarch Total RNA Miniprep Kit (New England Biolabs), according to manufacturer's instructions. Total RNA were sent in dry-ice package to Macrogen Europe B.V. Macrogen Europe BV constructed libraries using Illumina TruSeq stranded mRNA library preparation with Ribozero rRNA depletion.
disease
colorectal carcinoma
organism
Homo sapiens
organism
Homo sapiens
collection date
not collected
scientific_name
Homo sapiens
common name
human
organism part
colon
developmental stage
adult
genotype
AID-degron Ki-67
isolate
not applicable
cell_line
HCT116
cell_type
colorectal cancer cell
geographic location (country and/or sea)
not collected