GSM3776472: K562wt_NS; Homo sapiens; OTHER
NS-seq: DNA fractionation was performed using a 5-30% sucrose gradient to collect DNA fractions ranging from 0.5 to 2 kb. Five prime single-strand DNA ends were phosphorylated by T4 polynucleotide kinase (T4PK) (NEB, M0201S). After phenol/chloroform treatment to remove T4 PK, DNA was precipitated, resuspended and then treated with lambda-exonuclease (NEB, M0262S) to remove genomic DNA fragments that lacked the phosphorylated RNA primer. After RNAse treatment and DNA purification (Qiagen PCR purification kit, 28004), single stranded nascent strands were random-primed using the Klenow and DNA Prime Labeling System (Invitrogen, 18187013) TruSeq_Pair_End_Seq_Sample_Prep