A front limb of a quadruped. (The Random House College Dictionary, 1980)
Attributes by original data submitter
Sample
source_name
posterior forelimb bud
strain/background
Swiss Webster
genotype/variation
wild type
tissue
posterior forelimb bud
age
E10.5 (35 somites)
Sequenced DNA Library
library_strategy
ATAC-seq
library_source
GENOMIC
library_selection
other
library_construction_protocol
Limb buds were dissected, dissociated using 100ug/mL Liberase (Roche 05401119001) in PBS at 37 degrees for 6 minutes, and cells were counted. 5,000 cells per sample were lysed and immediately added to a transpose reaction for 30 min at 37C (Illumina FC-121-1030). After the transposase reaction was complete, the entire reaction was put into a PCR library amplification reaction. Barcoded primers, specific to the inserted transposon sequence, amplified accessible chromatin fragments. Fragments were amplified for 18 PCR cycles using NEBNext High Fidelity 2x PCR Master Mix (M0541S). Excess primers were removed from PCR reactions using AmpureXP beads (A63880) at a ratio of 0.8x beads.