organism: Escherichia coli str. K-12 substr. MG1655
description: Protocols: Full-length recombinant human DNMT3A (ab170408) and DNMT3B (ab170410) were purchased from Abcam. 100 ng unmethylated E. coli genomic DNA (D5016, Zymo research) was incubated at 37 °C with 500 ng of DNMT3A or DNMT3B and 160 µM S-adenosylmethionine (SAM, Cat. No. B9003S, NEB) in reaction buffer (50 mM Tris-HCI, 1 mM EDTA, 1 mM dithiothreitol, 5% glycerol, 100 µg/ml BSA) for 30, 120 min. DNA was purified using DNA Clean & Concentrator Kit (D4030, Zymo research) and processed for high throughput bisulfite sequencing. Bisulfite library was prepared using Pico Methyl-Seq Library Prep Kit (D5456, Zymo research) per manufacture's protocol. Briefly, methylated DNA and unmethylated E. coli DNA were treated with bisulfite conversion reagent at 98 °C for 8 min, then 54 °C for 60 min.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-05-15T04:03:52Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-10-04T11:38:29Z
attributes.tag: External Id
attributes.value: SAMEA5996553
attributes.tag: INSDC center name
attributes.value: CRUK-CI and Department of Chemistry, University of Cambridge
attributes.tag: INSDC first public
attributes.value: 2020-05-15T04:03:52Z
attributes.tag: INSDC last update
attributes.value: 2019-10-04T11:38:29Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8406:DNMT3A-Abcam-30min
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: sample name
attributes.value: E-MTAB-8406:DNMT3A-Abcam-30min
attributes.tag: scientific_name
attributes.value: Escherichia coli str. K-12 substr. MG1655
attributes.tag: strain
attributes.value: K-12
attributes.tag: substrain
attributes.value: MG1655