DNMT3B-Abcam-120min
Escherichia coli str. K-12 substr. MG1655
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.
ENA-FIRST-PUBLIC
2020-05-15T04:03:52Z
ENA-LAST-UPDATE
2019-10-04T11:38:29Z
External Id
SAMEA5996554
INSDC center name
CRUK-CI and Department of Chemistry, University of Cambridge
INSDC first public
2020-05-15T04:03:52Z
INSDC last update
2019-10-04T11:38:29Z
INSDC status
public
Submitter Id
E-MTAB-8406:DNMT3B-Abcam-120min
broker name
ArrayExpress
sample name
E-MTAB-8406:DNMT3B-Abcam-120min
scientific_name
Escherichia coli str. K-12 substr. MG1655
strain
K-12
substrain
MG1655