Bio-Rad CHEF Genomic DNA Plug Kits - Manual (Page 12)

LIT510 Rev E
Life Science
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Bio-Rad
Laboratories
and do not have to be dispersed by chemical or enzymatic
methods. The Grams Crystal Violet aids in the visualization
of bacteria.
Use the equations below to determine the cell
concentration:
For example: 300 bacteria in 5 squares = average of
60 bacteria/square x 25 (squares) x 20 (dilution factor, yeast
use 100 for dilution factor) x 10
4 = 3 x 10 8 bacteria per ml.
For 5 ml of plugs you need 5 ml x (5 x 10 8 ) cells final
concentration
÷
(3 x 10 8 ) actual cells concentration = 8.33 ml
of cell suspension is required.
For yeast use 6 x 10 8 cells per ml for the final
concentration (see page 9).
Cells Counted
Number of Squares
= Average Cells per Square
Average Cells per square × 25 Squares × Dilution Factor × 10 4 = Cells per ml
desired cell concentration
actual cell concentration
× ml of plugs to be made = ml of cell suspension
to use.
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LIT510E 2/11/99 3:47 PM Page 18