title: GSM5929393: Arabidopsis_cauline - IgG antibody IP-seq; Arabidopsis thaliana; OTHER
library_construction_protocol: Cauline leaves were collected from three separate flowering col-0 Arabidopsis thaliana plants and combined together. Two biological replicates were obtained from plants grown 3 months apart from each other. Approximately 1 g of Cauline leaves was combined with 1.2 mL of Buffer A supplemented with 1% sodium dodecyl sulfate (Sigma L3771), 0.5% polyvinylpyrrolidone (Sigma PVP40) and fresh 1% 2-Mercaptoethanol (Sigma M3148) in a Dounce homogenizer. Tissue was ground using a “B” pestle until homogenous. The lysate was transferred in 200 μl aliquots to 1.5 mL microfuge tubes and all samples were mixed with 3 μl Proteinase K and incubated for 1 hour at 50°C. The DNA was purified by adding 200 μl (1:1) of phenol:chloroform:isoamyl alcohol 25:24:1 (saturated with 10mM Tris, pH 8.0, 1mM EDTA) then mixed by vigorous tube inversions and incubated for 10 min at RT.