Since the denaturing conditions in TTGE span a wider range than CDGE, several fragments with different melting behavior, that are analyzed on separate gels by CDGE, can be analyzed on the same TTGE gel. This is illustrated in Figure 3, where exons 59 and 11 of the TP53 gene are analyzed. Using two different denaturing conditions and the same temperature ramp allowed all of the exons to be analyzed at one time.
These examples demonstrate that the TTGE technique is more robust and flexible than either DGGE or CDGE in mutation screening and represents a new development with advantages for mutation screening in a diagnostic setting.
With TTGE, the high reproducibility in casting a constant denaturant gel and the rapid and high throughput of the analyses obtained by CDGE are achieved. The problems in determining the exact denaturing conditions for fragments with only one melting domain are eliminated. The focusing of bands obtained by DGGE seems to be achieved, and since the denaturing conditions in TTGE span a wider range, several fragments with different melting behavior can be analyzed on the same gel.
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