organism: Homo sapiens
description: Protocols: All lung cancer blood samples were obtained from patients with adenocarcinoma and control blood samples were from healthy volunteers. All blood from lung cancer patients and controls (∼5 ml), was collected in tubes containing EDTA as anti-coagulant. Centrifuge blood samples for 15 min at 1500 x g, and then transfer plasma to clean tubes. The plasma was aliquoted in ~1 ml fractions to avoid multiple freeze-thaw times and stored at -80℃ until DNA extraction. The cfDNA was extracted from 1ml plasma using QIAamp Circulating Nucleic Acid kit (Qiagen, 55114) according to the manufacturer's protocol. Qubit (Invitrogen) was used to analysis the concentration of cfDNA in the plasma. The distribution of cfDNA size was estimated by the Agilent Bioanalyzer 2100. The obtained cfDNA was performed for end repair, dA overhanging, Illumina sequencing adapter ligation to get original library. Then, the library was denatured at 95°C for 10 min, immediately ten minutes on ice, then incubated with 5-Methylcytosine (5-mC) Monoclonal Antibody (Epigentek, A-1014) that was bound on Magna ChIP Protein A+G Magnetic beads (Millipore, 16-663) for 4°C overnight, and decrosslinked subsequently. The products were amplified using Q5 High-Fidelity DNA Polymerase (NEB, M0491). The PCR reactions were then purified using AMPure XP beads (Beckman). Amplificated libraries were detected DNA fragmentation size by Agilent Bioanalyzer 2100 (Agilent Technologies) and determined concentration with the Qubit assay (Invitrogen).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-03-31T17:01:55Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-08-22T15:55:53Z
attributes.tag: External Id
attributes.value: SAMEA4852661
attributes.tag: INSDC center name
attributes.value: SHANGHAI JIAO TONG UNIVERSITY
attributes.tag: INSDC first public
attributes.value: 2019-03-31T17:01:55Z
attributes.tag: INSDC last update
attributes.value: 2018-08-22T15:55:53Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7163:Sample 8_5-mC
attributes.tag: age
attributes.value: not available
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: normal
attributes.tag: individual
attributes.value: control3
attributes.tag: organism part
attributes.value: blood plasma
attributes.tag: sample name
attributes.value: E-MTAB-7163:Sample 8_5-mC
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: not available