Hoefer HE99X - Manual (Page 21)

Agarose gel electrophoresis notes
Agarose gel electrophoresis can be used to
separate DNA fragments down to 0.1 kb or
less. Polyacrylamide gels are typically used for
fragments smaller than 1 kb.
DNA mobility
The suggested agarose concentration for
separating fragments of various sizes is listed
below. Other factors affecting separation results
include the selected running buffer, the voltage
setting, the temperature, and the presence of
ethidium bromide.
Agarose concentrations for separating
DNA fragments of various sizes
effective range of resolution
agarose (%) of linear DNA fragments (kb) †
0.5 1 to 30
0.7 0.8 to 12
1.0 0.5 to 10
1.2 0.4 to 7
1.5 0.2 to 3
† Current Protocols in Molecular Biology, p 2.5.2 (1993).
A common standard is a Hind III digest of
lambda phage, which gives eight fragments rang-
ing in size from 0.1 to 23 kb. The bands are well
resolved when run 2 hours on a 20 cm long 1%
agarose gel in 0.5X TBE buffer at 150 V.
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