title: GSM6601085: RNAseq, CHD6_Control_shScr1; Homo sapiens; RNA-Seq
library_construction_protocol: RNA-Seq: Total RNA was isolated from cells using the RNA MiniPrep kit (ZYMO Research). ChIP-Seq: Chromatin DNA in ChIP-Seq samples was purified by using columns provided in Magna ChIP A/G Kit (Millipore Cat.17-10085). MNase-Seq: Nucleosome DNA was extracted using phenol:chloroform:isoamyl for high-throughput sequencing. RNA-Seq: 1. Take a certain amount of total RNA samples, and use oligo dT beads to enrich mRNA with poly A tail; 2. mRNA molecules were fragmented into small pieces; 3. The fragmented mRNA was synthesized into first strand cDNA using random primers; 4. The second strand cDNA was synthesized with dUTP instead of dTTP; 5. The synthesized cDNA was subjected to end-repair and 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments; 6. Digest the U-labeled second-strand template with Uracil-DNA-Glycosylase (UDG) and perform PCR amplification; 7. Library quality control; 8. Library circularization; 9. The library was amplified to make DNA nanoball (DNB); 10. Sequencing on DNBSEQ (DNBSEQ Technology) platform. ChIP-Seq and MNase-Seq: (1) Take IPed-DNA and Input DNA for sample qualification; (2) DNA-end repair, 3'-dA overhang and adaptor ligation; (3) PCR amplification and size selection; (4) Library quality control; (5) Library circularization;(6) The library was amplified to make DNA nanoball (DNB); (7) Sequencing on DNBSEQ (DNBSEQ Technology) platform.