High performance capillary electrophoresis - T.E.A.M.

Modes
CGE can be altered by the addition of chiral selectors, ionpairing
reagents, or another complexing agent (such as
ethidium bromide for DNA and SDS for proteins). These
species can be covalently bound to the gel or simply added
to the running buffer.
3.3.1 Applications of CGE
CGE has been employed for both molecular biology and
protein chemistry applications. The former includes oligonucleotide
purity analysis, anti-sense gene therapy, DNA
sequencing, PCR product analysis, and DNA forensics. The
latter includes native and SDS-complexed protein separations.
Separation of double-stranded DNA restriction fragments,
produced by the enzymatic digest of large DNA, have also
been successful. Generally, the more open structure of linear
or minimally crosslinked gels are better suited for these large
molecules, which can range in size from a few base pairs to
more than 10 6 base pairs. The separation of a 1-kbp ladder,
for example, is shown in figure 43.
Figure 43
CGE of 1 kbp ladder using minimally
crosslinked polyacryl amide 22
Conditions: Bis-crosslinked polyacrylamide
(3 % T, 0.5 % C), 100 mM Trisborate,
pH 8.3, E = 250 V/cm,
i = 12.5 mA, l = 30 cm, L = 40 cm,
id = 75 mm, l = 260 nm,
polyacrylamide coated capillary
1 75
2 142
3 154
4 200
5 220
6 298
7 344
8 394
9 506
10 516
11 1018
12 1635
13 2036
1
bp
14
15
16
17
18
19
20
21
22
23
2 3 4 5 6
3054
4072
5090
6108
7126
8144
9162
10180
11198
12216
bp
7 8 9 10
10 15 20
Time [min]
11
12
13
15
14 16
17
18
19
20
21
22 23
73