PMC:6218659 / 83854-85037 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/6218659","sourcedb":"PMC","sourceid":"6218659","source_url":"https://www.ncbi.nlm.nih.gov/pmc/6218659","text":"Macromolecular incorporation assay\nMtb H37Rv ATCC27294 was grown to OD650nm of 0.6 in 7H9/ADC/Tw was treated at 37 °C under constant agitation with 20 μCi/mL of D[-6-3H]-glucosamine (American Radiolabeled Chemicals, Inc. 40 Ci/mmol) for 2 h followed by addition of compound (TUN-8,8, tunicamycin, meropenem/clavulanate or DMSO as vehicle control) at 1X or 10X MIC concentrations. MIC concentrations for controls were as follows: D-cycloserine 29 μM, meropenem 2 μM with clavulanate used at a fixed concentration of 100 μM, tunicamycin 1.1 μM. After 24 h and 48 h incubation, 100 μl of culture was precipitated with 100 μl of 20% TCA in a round bottom 96 well polystyrene (Nunclon) plate. Precipitates were aspirated with a cell harvester (Perkin Elmer) and transferred to 96 well filtermat. The precipitates on the GF/C filtermats (Perkin Elmer) were washed twice with 200 μl 10% TCA, and subsequently three times with 200 μl of 70% ethanol. The filtermat was removed, dried overnight and then soaked with 5 ml of Betaplate scintillation cocktail (Perkin Elmer) followed by mounting in a filtermat holder. It was counted by Microbeta2 microplate scintillation counter (Perkin Elmer).","divisions":[{"label":"Title","span":{"begin":0,"end":34}}],"tracks":[]}