Bio-Rad CHEF Genomic DNA Plug Kits - Manual (Page 10)

9.2 Agarose Concentrations
Various agarose plug percentages are reported in the
literature. The most common final concentration is 0.5%, but
the softness of these plugs makes them difficult to keep intact
and handle. We have found that 0.75% to 1% agarose greatly
enhances the strength of the plug without interfering with the
migration of the DNA out of the plug. These kits contain a 2%
CleanCut agarose solution, which when mixed with differing
amounts of Cell Suspension Buffer, will give agarose
concentrations ranging from 0.5 to 1%. This allows the option
of using a different final concentration of agarose if desired.
* Vol. is the total volume of plugs to be made at 75 µl/plug, i.e. for 5 ml
of plugs at 0.75% final agarose concentration, add (0.375)5 ml = 1.9 ml
of 2% agarose solution to (0.625)5 ml = 3.4 ml of cell suspension.
Desired Agarose
Concentration
CleanCut Agarose
2%*
1% (0.5)vol. = ____
0.75%
0.5%
(0.375)vol. = ____
(0.25)vol. = ____
Cell Suspension
Buffer*
(0.5)vol. = ____
(0.625)vol. = ____
(0.75)vol. = ____
15
Section 9
Appendix
9.1 Solutions
PMSF, 100 mM Add 0.174 g to 10 ml of 100%
(phenylmethanesulfonyl isopropanol. Store at room tem-
fluoride) perature.
A stock solution of 100 mM in
100% isopropanol is stable at
room temp for 1 year. PMSF is
not stable in aqueous solutions.
Activity lasts approximately 30 to
50 min at room temperature at pH
7.5-8.
LB Per liter:
10 g Bacto Tryptone
5 g Bacto Yeast Extract
10 g NaCl
pH 7
YPD Per liter:
20 g Bacto Peptone
10 g Bacto Yeast Extract
20 g dextrose (glucose)
14
LIT510E 2/11/99 3:46 PM Page 14