title: GSM4085154: TMU38_lncRNA; Triticum urartu; RNA-Seq
library_construction_protocol: The leaves were then transferred to liquid nitrogen and ground into a fine powder. The following experiment was carried out using the native ChIP protocol. The chromatin was digested into single nucleosomes using 0.02 U/μl DNase I (RNase-free) at 37°C. Dynabeads Protein A (ThermoFisher, Cat#:10001D) was used for binding the anti-CENH3 antibody. After washing the beads incubated with the antibody and chromatin fragments, the beads were eluted twice at 65°C for 15 min using elution buffer containing 50 mM Tris-HCl (pH = 7.5), 10 mM EDTA, 200 mM NaCl and 1% SDS. The eluted fractions were combined, and Proteinase K (Invitrogen, Cat#: 25530-49) was added to relieve cross-linking at 37°C overnight after which the RNA was extracted using TRIzol reagent, and the residual DNA was removed using RNase-free DNase I (NEB, Cat#: M0303S). The cDNA libraries of RIP samples were prepared using the standard manual provided in the NEB Next Ultra RNA Library Prep Kit for Illumina (NEB, Cat#: E7530S).