Download e-book for kindle: Current Protocols in Molecular Biology by Frederick M. Ausubel

By Frederick M. Ausubel

ISBN-10: 047150338X

ISBN-13: 9780471503385

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6 Copyright © 2002 by John Wiley & Sons, Inc. 2 × 108 cells/ml. 6. Any of the single colonies may be saved for further use. Store plates at 4°C wrapped in parafilm or in the plastic sleeve in which the plates were supplied. Commentary Titering by serial dilutions is a good way to determine the number of any kind of living organism present in a suspension. 11). It is sometimes useful to use smaller factors of dilution. Mixing 50 µl of the culture into 5 ml medium will give dilutions of 100×. Mixing 100 µl into 900 µl will give dilutions of 10×.

2 Copyright © 2002 by John Wiley & Sons, Inc. 1 Supplement 59 With a Spectrophotometer The concentration of cells in a culture can also be determined with a spectrophotometer by measuring the amount of 600-nm light scattered by the culture. The level of absorbance (A) at 600 nm will depend on the distance between the cuvette and the detector and will vary among spectrophotometers, often by a factor of 2. 3). If the culture is visibly turbid, also measure a 10-fold dilution of it. 1 OD unit is roughly equivalent to 1 × 108 cells/ml.

Identification and characterization of recD, a gene affecting plasmid maintenance and recombination in Escherichia coli. J. Bacteriol. 167:594-603. Bolivar, F. W. 1977. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95-113. E. 1976. The construction in vitro of transducing derivatives of phage lambda. Mol. Gen. Genet. 146:199-207. W. and Roulland-Dussoix, D. 1969. A complementation analysis of the restriction and modification of DNA in Escherichia coli.

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Current Protocols in Molecular Biology by Frederick M. Ausubel

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