Handbook of Size Exclusion Chromatography and Related ...

Table 1 Exclusion Limits of TSKgel SW and PW Columns for RNA and Double-
Stranded DNA Fragments a
6 COLUMNS
Two types of columns have been employed in the SEC of nucleic acids:
chemically bonded porous silica columns and hydrophilic resin columns.
Among them, TSKgel SW and PW columns have been well accepted. They are
available in different pore sizes, and each has a different separation range. The
exclusion limits for RNA and double-stranded DNA fragment are listed in
Table 1. A sample of a certain molecular weight can be in general separated on
different columns. However, the resolution depends on the column employed.
For example, in the separation of HaeIII-cleaved plasmid pBR322, the best
separation is obtained for base pairs of 7–21, 51–104, 123–267, and 434–587
on G2000SW, G3000SW, G4000SW, and G5000PW, respectively. Therefore, it
is very important to select the best column depending on the molecular weights
Table 2 Best Columns for the Separation of RNA
Molecular weight range Best column
,60,000 G2000SW or G3000SW
60,000–120,000 G3000SW
120,000–1,200,000 G4000SW
1,200,000–10,000,000 G5000PW
Source: Ref. 8.
Exclusion limit (molecular weight)
Column RNA Double-stranded DNA fragment
G2000SW 70,000 50,000 (70) b
G3000SW 150,000 100,000 (150)
G4000SW 1,500,000 300,000 (500)
G5000PW .5,000,000 1,000,000 (1,500)
G6000PW — c
5,000,000 (7,000)
DNA-PW — c
5,000,000 (7,000)
a
In 0.1 M phosphate buffer (pH7.0) containing 0.1 M sodium chloride and 1mM EDTA.
b
Values in parentheses are the exclusion limits in base pairs.
c
Not determined.
Source: Refs. 8 and 12.
© 2004 by Marcel Dekker, Inc.