Handbook of Size Exclusion Chromatography and Related ...

Although there are avariety of species in tRNA, their molecular weights
are in anarrow range, approximately 25,000–30,000. Therefore, it is rather
difficult to separate different species of tRNA by SEC. Single peaks are usually
observed in SEC of tRNA samples even if they contain many species. Only one
example of the separation of tRNA species has been reported. Two species,
tyrosine-specific and N-formylmethionyl-specific tRNAs, were separated on a
MicroPak TSK 3000SW column (30 cm 7.5 mm inner diameter, ID),
although only partially (1). However, it is easy to separate tRNA from other
types of RNA such as rRNA, as exemplified in Fig. 1. tRNA was separated
from rRNA on a TSKgel G3000SW two-column system (each column
60 cm 7.5 mm ID).
Separation of different species of rRNA is also easy. Figure 2shows an
example of the separation of 5S, 16S, and 23S rRNAs, whose molecular weights
are approximately 39,000, 560,000, and 1,100,000; they were separated well on a
TSKgel G4000SW two-column system (each column 60 cm 7.5 mm ID) in
Figure 1 Separation of total E. coli RNAs containing 4s tRNA and 5S, 16S, and 23S
rRNAs obtained on a TSKgel G3000SW two-column system (each column
60 cm 7.5 mm ID) in 0.1 M phosphate buffer (pH 7.0) containing 0.1 M sodium
chloride and 1 mM EDTA at a flow rate of 1 mL/min. (From Ref. 9.)
© 2004 by Marcel Dekker, Inc.