De novo fatty acid and mycolic acid biosyntheses were followed by labeling 5 ml culture aliquots with 1 µCi/ml [1-14C]-acetate (specific activity: 55.3 mCi/mmol; Perkin Elmer) for 1 h at 37°C. Fatty acid and mycolic acid methyl esters were extracted from samples containing equivalent amounts of bacteria as described by [32] (link). The resulting solution of FAMEs and MAMEs was assayed for radioactivity in a Beckman liquid scintillation counter and then subjected to TLC using silica gel plates (5735 silica gel 60F254; Merck). Samples were normalized by culture OD or total cpm and developed in hexane∶ethyl acetate (9∶1, v/v). Autoradiograms were produced by overnight exposure to Kodak X-Omat AR film to reveal 14C-labelled FAMEs and MAMEs.
Liquid scintillation counter
Manufactured by Merck Group
The Liquid Scintillation Counter is an analytical instrument used to detect and measure the presence of radioactive materials in liquid samples. It functions by mixing the liquid sample with a scintillation cocktail, which converts the energy of the radioactive emissions into light pulses that are then detected and quantified by the instrument.
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Lab products found in correlation
2 protocols using liquid scintillation counter
1
Labeling Mycolic Acid Biosynthesis
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De novo fatty acid and mycolic acid biosyntheses were followed by labeling 5 ml culture aliquots with 1 µCi/ml [1-14C]-acetate (specific activity: 55.3 mCi/mmol; Perkin Elmer) for 1 h at 37°C. Fatty acid and mycolic acid methyl esters were extracted from samples containing equivalent amounts of bacteria as described by [32] (link). The resulting solution of FAMEs and MAMEs was assayed for radioactivity in a Beckman liquid scintillation counter and then subjected to TLC using silica gel plates (5735 silica gel 60F254; Merck). Samples were normalized by culture OD or total cpm and developed in hexane∶ethyl acetate (9∶1, v/v). Autoradiograms were produced by overnight exposure to Kodak X-Omat AR film to reveal 14C-labelled FAMEs and MAMEs.
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De novo fatty acid and mycolic acid biosyntheses were followed by labeling 5 ml culture aliquots with 1 µCi/ml [1-14C]-acetate (specific activity: 55.3 mCi/mmol; Perkin Elmer) for 1 h at 37°C. Fatty acid and mycolic acid methyl esters were extracted from samples containing equivalent amounts of bacteria as described by [32] (link). The resulting solution of FAMEs and MAMEs was assayed for radioactivity in a Beckman liquid scintillation counter and then subjected to TLC using silica gel plates (5735 silica gel 60F254; Merck). Samples were normalized by culture OD or total cpm and developed in hexane∶ethyl acetate (9∶1, v/v). Autoradiograms were produced by overnight exposure to Kodak X-Omat AR film to reveal 14C-labelled FAMEs and MAMEs.
2
Conditional M. smegmatis Fatty Acid Analysis
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